CN116407582B

CN116407582B - Compound preparation for treating RA and multi-organ complications thereof and preparation method thereof

Info

Publication number: CN116407582B
Application number: CN202111641995.6A
Authority: CN
Inventors: 梁倩倩; 王拥军; 施杞; 徐浩; 李雪菲; 陈涛; 李强; 常君丽
Original assignee: Longhua Hospital Affiliated to Shanghai University of TCM
Current assignee: Longhua Hospital Affiliated to Shanghai University of TCM
Priority date: 2021-12-29
Filing date: 2021-12-29
Publication date: 2024-03-29
Anticipated expiration: 2041-12-29
Also published as: CN116407582A

Abstract

The invention provides a composition for treating Rheumatoid Arthritis (RA) and complications of multiple organs thereof. Specifically, the invention provides a composition, which comprises raw medicinal materials or extracts thereof, wherein the raw medicinal materials comprise: radix astragali, radix Angelicae sinensis, bupleuri radix, radix Gentianae Marcrophyllae, rhizoma corydalis, radix Cyathulae, herba Epimedii, and herba et Gemma Agrimoniae. The composition of the invention can improve the complications of multiple organs caused by RA, obviously reduce inflammation and improve the treatment effect.

Description

Compound preparation for treating RA and multi-organ complications thereof and preparation method thereof

Technical Field

The invention relates to the field of traditional Chinese medicine treatment, in particular to a traditional Chinese medicine composition for treating rheumatoid arthritis and multi-organ complications caused by the rheumatoid arthritis, an extracting agent and a compound preparation of the traditional Chinese medicine composition.

Background

Rheumatoid arthritis (Rheumatoid Arthritis, RA) is an autoimmune disease characterized by extensive persistent arthromeningitis and symmetrical, destructive arthropathy. The traditional Chinese medicine composition is good for young and strong, low in alleviation rate, high in disability rate and high in treatment cost, and occupies 0.5% -1.0% of the world population.

Although arthritis is a major symptom of RA, it can cause systemic multi-organ damage and severely affect lung, heart, liver, kidney, spleen functions in the progression of the disease and with the use of long-term antirheumatic drugs.

At present, clinical treatment is not clearly guided, and a safe and effective treatment method for multi-organ complications caused by RA is also lacking, and the most commonly used medicines for treating RA are acknowledged to be antirheumatic medicines and nonsteroidal anti-inflammatory medicines, but the curative effect is very low, the aim of treating RA and the multi-organ complications cannot be achieved, and the risk of damaging liver and kidney functions is possibly caused after long-term use.

Therefore, the development of safer and more effective medicaments for treating RA and complications thereof has become a great problem to be researched and solved in the aspects of saving the lives of patients, relieving clinical symptoms and improving the quality of life.

Disclosure of Invention

The invention aims to provide a traditional Chinese medicine composition, an extracting agent and a pharmaceutical preparation thereof for effectively treating rheumatoid arthritis and complications of multiple organs.

In a first aspect of the present invention, there is provided a Chinese medicinal composition comprising crude drugs or extracts thereof, wherein the crude drugs comprise: radix astragali, radix Angelicae sinensis, bupleuri radix, radix Gentianae Marcrophyllae, rhizoma corydalis, radix Cyathulae, herba Epimedii, and herba et Gemma Agrimoniae.

In another preferred embodiment, the crude drug comprises fresh drug, partially dried drug, or a combination thereof.

In another preferred example, when the composition comprises the raw medicinal materials, parts by weight thereof are calculated based on dry weight of the respective raw medicinal materials.

In another preferred embodiment, when the composition comprises an extract of a crude drug, the compatibility of the extract is calculated as dry weight of the corresponding crude drug.

In another preferred embodiment, the extract comprises: the aqueous extract, the alcoholic extract, the aqueous solvent extract, the carbon dioxide supercritical extract or the active ingredient, or a combination thereof, is preferably an aqueous extract, an alcoholic extract.

In another preferred embodiment, the alcohol comprises: C1-C6 alcohols such as methanol, ethanol, propanol, or combinations thereof.

In another preferred embodiment, the active ingredient comprises a single ingredient or a mixture of ingredients.

In another preferred embodiment, the Chinese medicinal composition comprises:

2-70 parts by weight (preferably 4-50 parts by weight, more preferably 10-50 parts by weight, for example 10, 15, 18, 20, 25, 30, 40, 50) of Astragalus mongholicus;

2-50 parts by weight of angelica (preferably 4-30 parts by weight, more preferably 10-30 parts by weight, e.g. 10, 12, 15, 20, 25);

2-50 parts by weight (preferably 4-30 parts by weight, more preferably 5-20 parts by weight, e.g. 5, 9, 10, 15, 20) of bupleurum;

2-50 parts by weight of gentiana macrophylla (preferably 4-30 parts by weight, more preferably 5-20 parts by weight, e.g. 5, 9, 10, 15, 20);

2-50 parts by weight (preferably 4-30 parts by weight, more preferably 5-20 parts by weight, e.g. 5, 6, 8, 9, 10, 15, 20) of corydalis tuber;

2-50 parts by weight (preferably 4-30 parts by weight, more preferably 10-30 parts by weight, for example 10, 12, 15, 20, 25) of cyathula root;

2-50 parts by weight (preferably 4-30 parts by weight, more preferably 10-30 parts by weight, for example 10, 12, 15, 20, 25) of herba Epimedii;

2-50 parts by weight (preferably 4-30 parts by weight, more preferably 10-30 parts by weight, for example 10, 12, 15, 20, 25) of hairyvein agrimony.

In another preferred embodiment, the Chinese medicinal composition comprises:

4-50 parts of raw astragalus mongholicus;

4-30 parts of Chinese angelica;

4-30 parts of bupleurum;

4-30 parts of gentiana macrophylla;

4-30 parts of rhizoma corydalis;

4-30 parts of radix cyathulae;

4-30 parts of herba epimedii;

4-30 parts of hairyvein agrimony.

In another preferred embodiment, the Chinese medicinal composition comprises:

10-50 parts of raw astragalus mongholicus;

10-30 parts of Chinese angelica;

5-20 parts of bupleurum;

5-20 parts of gentiana macrophylla;

5-20 parts of rhizoma corydalis;

10-30 parts of radix cyathulae;

10-30 parts of herba epimedii;

10-30 parts of hairyvein agrimony.

In another preferred example, the weight ratio of the Chinese medicinal composition, namely, angelica sinensis, radix bupleuri, gentiana macrophylla, medicinal cyathula root, epimedium herb and hairyvein agrimony is (0.7-1.2): (0.7-1.2): (0.7-1.2): (0.7-1.2): (0.7-1.2): (0.7-1.2), preferably 1: (0.75-1): (0.75-1): 1:1:1.

in another preferred example, in the traditional Chinese medicine composition, the weight ratio of raw astragalus, angelica, radix bupleuri, gentiana macrophylla, rhizoma corydalis, medicinal cyathula root, epimedium herb and hairyvein agrimony is (1.5-2.5): (0.7-1.2): (0.7-1.2): (0.7-1.2): (0.5-1): (0.7-1.2): (0.7-1.2): (0.7-1.2), preferably 2:1: (0.75-1): (0.75-1): 0.5:1:1:1.

in another preferred embodiment, the Chinese medicinal composition comprises: 18 g of raw astragalus, 12 g of Chinese angelica, 9 g of bupleurum, 9 g of gentiana macrophylla, 6g of corydalis tuber, 12 g of medicinal cyathula root, 12 g of epimedium, and 12 g of hairyvein agrimony.

In another preferred embodiment, the Chinese medicinal composition comprises: 30 g of raw astragalus, 12 g of Chinese angelica, 12 g of radix bupleuri, 12 g of gentiana macrophylla, 9 g of rhizoma corydalis, 12 g of medicinal cyathula root, 12 g of herba epimedii and 12 g of hairyvein agrimony.

In a second aspect of the present invention, there is provided a Chinese medicinal composition extractant prepared by extraction of the Chinese medicinal composition according to the first aspect of the present invention, wherein the extraction solvent of the extractant is selected from the group consisting of: water and ethanol.

In another preferred embodiment, the ratio of the Chinese medicinal composition to the solvent for extraction is 1:6-20, such as 8, 10, 14.

In another preferred embodiment, the extractant includes an unregulated (or unchanged) concentration of extractant, a concentrated extractant, a diluted extractant.

In another preferred embodiment, the extractant is a decoction or an extract of the traditional Chinese medicine composition.

In another preferred embodiment, the extractant is a concentrated decoction or extract.

In another preferred embodiment, the relative density of the extract is 1-1.4, preferably 1.05-1.35 or 1.3-1.4.

In a third aspect of the present invention, there is provided a method of preparing a Chinese medicinal composition according to the first aspect of the present invention or an extractant of a Chinese medicinal composition according to the second aspect of the present invention,

i) The method comprises the following steps:

a1 Providing a crude drug, the crude drug being as defined in claim 1;

b1 Mixing the raw materials with water, decocting, and filtering to obtain decoction A;

c1 Concentrating to obtain extract A;

d1 Mixing the extract A obtained in the step c 1) with alcohol, standing, and concentrating for the second time to obtain an extract B;

alternatively, ii) comprises the steps of:

a2 Providing a crude drug, the crude drug being as defined in claim 1;

b2 Mixing the raw materials with alcohol, decocting, and filtering to obtain decoction B;

c2 Concentrating to obtain extract C;

alternatively, iii) comprises the steps of:

a3 Providing a crude drug, the crude drug being as defined in claim 1;

b3 Decocting the raw materials, adding alcohol into the first decoction, adding water into the second decoction, and filtering to obtain decoction C;

c3 Concentrating to obtain extract D.

In another preferred embodiment, the alcohol is selected from the group consisting of: water, methanol, ethanol, propanol, or combinations thereof, ethanol being preferred.

In another preferred embodiment, the alcohol is 50-90% ethanol by volume.

In another preferred embodiment, the temperature of the decoction is 60-100deg.C.

In another preferred embodiment, in the step b 1), the mass ratio of the raw material medicine to water is 8-14.

In another preferred embodiment, in step b 1), the said decoction is 2-3 times, each independently for a time of 0.5-5 hours, preferably 1-3 hours.

In another preferred embodiment, in step c 1), the relative density of the extract A is from 1 to 1.5, preferably from 1.05 to 1.35.

In another preferred embodiment, the step d 1) includes: mixing the extract A obtained in the step c 1) with ethanol to ensure that the alcohol concentration is 50-80%, standing, and concentrating the supernatant twice to obtain an extract B.

In another preferred embodiment, the relative density of the extract B is 1-1.5, preferably 1.3-1.4.

In another preferred embodiment, in step b 2), the mass ratio of the crude drug to the alcohol is 8-14, preferably 8-10.

In another preferred embodiment, in step b 2), the decoction is 2-3 times, each independently for a period of 0.5-5 hours, preferably 1-3 hours.

In another preferred embodiment, in step C2), the relative density of the extract C is between 1 and 1.5, preferably between 1.05 and 1.4.

In another preferred embodiment, in step b 3), the alcohol is 50-90% ethanol by volume fraction.

In another preferred embodiment, in step b 3), the mass ratio of the crude drug to the alcohol is 8-14, preferably 8-10; the mass ratio of the raw medicinal materials to water is 6-14, preferably 6-10.

In another preferred embodiment, in step b 3), the time of the decoction is each independently from 0.5 to 5 hours, preferably from 1 to 3 hours.

In another preferred embodiment, in step c 3), the relative density of the extract D is 1-1.5, preferably 1.3-1.4.

In another preferred embodiment, the method further comprises a post-treatment step of drying under reduced pressure and pulverizing to obtain extract powder.

In a fourth aspect of the invention, there is provided a pharmaceutical formulation comprising:

i) The traditional Chinese medicine composition according to the first aspect of the invention or the traditional Chinese medicine composition extractant according to the second aspect of the invention;

ii) other pharmaceutically acceptable adjuvants, carriers or excipients.

In another preferred embodiment, the pharmaceutical formulation is a traditional Chinese medicine formulation.

In another preferred embodiment, the formulation is an oral formulation or a non-oral formulation.

In another preferred embodiment, the formulation comprises a liquid formulation, a semi-solid formulation, or a solid formulation.

In another preferred embodiment, the formulation comprises: powder, granule, capsule, mixture, powder, injection, tincture, oral liquid, tablet, buccal tablet, or dripping pill.

In another preferred embodiment, the preparation method of the preparation comprises the following steps:

the extract of the second aspect of the invention is post-treated to obtain extract powder, and the extract powder is added with medicinal auxiliary materials, evenly mixed, granulated and tabletted to prepare tablets with 0.1-1g of each tablet.

In a fifth aspect of the present invention, there is provided the use of a Chinese medicinal composition according to the first aspect of the present invention or a Chinese medicinal composition extractant according to the second aspect of the present invention or a pharmaceutical formulation according to the third aspect of the present invention for the manufacture of a medicament and/or health care product for the prevention and/or treatment of rheumatoid arthritis and/or multi-visceral complications thereof.

In another preferred embodiment, the rheumatoid arthritis is an autoimmune disease resulting from stimulation with an antigen, wherein the antigen comprises lipopolysaccharide, a virus, a bacterial toxin.

In another preferred embodiment, the rheumatoid arthritis is an autoimmune disease resulting from lipopolysaccharide stimulation.

In another preferred embodiment, the multi-organ complication is a complication of an organ selected from the group consisting of rheumatoid arthritis: lung, heart, liver, kidney, spleen, or a combination thereof.

In another preferred embodiment, the multi-organ complication is selected from the group consisting of: interstitial pneumonia, myocardial hypertrophy, autoimmune liver disease, chronic nephritis, splenomegaly, or a combination thereof.

In another preferred embodiment, the prevention and/or treatment of rheumatoid arthritis and/or a multi-visceral complication resulting therefrom comprises an improvement in one or more indicators selected from the group consisting of:

a) Reducing inflammatory cell infiltration and/or pulmonary fibrosis in the lung;

b) Improving myocardial hypertrophy and/or cardiac fibrosis;

c) Reducing hepatic vascular inflammation and/or hepatic fibrosis;

d) Reducing inflammation, fibrosis and/or glomerulopathy of the renal interstitial tissue;

e) Alleviating splenomegaly.

In a sixth aspect of the invention, there is provided a method of treating RA and its multiple organ complications, comprising the steps of: administering to a subject in need thereof a pharmaceutically effective amount of the composition of the first aspect or the pharmaceutical formulation of the third aspect.

In another preferred embodiment, the subject is a rheumatoid arthritis patient or a rheumatoid arthritis and complications thereof.

In another preferred embodiment, the multi-visceral complications include injury to the lung, heart, liver, kidney, spleen.

It is understood that within the scope of the present invention, the above-described technical features of the present invention and technical features specifically described below (e.g., in the examples) may be combined with each other to constitute new or preferred technical solutions. And are limited to a space, and are not described in detail herein.

Drawings

FIG. 1 is a graph of lung HE and Masson staining measurements in animal experiments.

FIG. 2 is a graph of results of heart HE and Masson staining measurements in animal experiments.

FIG. 3 is a graph of liver HE and Masson staining measurements in animal experiments.

FIG. 4 is a graph of the results of the detection of renal HE and Masson staining in animal experiments.

FIG. 5 is a graph showing the results of detection of spleen HE staining in animal experiments.

Detailed Description

The present inventors have conducted extensive and intensive studies and have developed, for the first time, a Chinese medicinal composition for treating complications of multiple organs caused by rheumatoid arthritis. Specifically, the composition of the invention is a 'dirty arthralgia formula', which comprises astragalus root, angelica, bupleurum, gentiana macrophylla, corydalis tuber, medicinal cyathula root, epimedium and hairyvein agrimony, and has the functions of tonifying qi, activating blood, dispelling wind, removing dampness, dredging arthralgia and relieving pain. The composition can prevent, treat and/or alleviate multi-organ complications generated by arthritic patients, alleviate injuries of lungs, hearts, livers, kidneys and spleens, and is very suitable for preparing pharmaceutical compositions and/or health-care product compositions for preventing, treating and/or alleviating rheumatoid arthritis and multi-organ complications caused by the rheumatoid arthritis. The present invention has been completed on the basis of this finding.

Terminology

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

As used herein, the terms "comprising," "including," and "containing" are used interchangeably, and include not only closed-form definitions, but also semi-closed-form and open-form definitions. In other words, the term includes "consisting of … …", "consisting essentially of … …".

As used herein, the term "relative density" refers to the ratio of the density of a substance to the density of a reference substance, which in the present invention is the density of water at 4 ℃ (used as 1), at the respective prescribed conditions, and the relative density is the ratio of the density of another substance (e.g., extract) to the density of water at 4 ℃.

As used herein, the term "pharmaceutically acceptable adjuvant, carrier or vehicle" component refers to a substance that is suitable for use in humans and/or animals without undue adverse side effects (such as toxicity, irritation, and allergic response), commensurate with a reasonable benefit/risk ratio.

As used herein, the term "effective amount" refers to an amount that is functional or active in and acceptable to a human and/or animal. It will be appreciated by those of ordinary skill in the art that the "effective amount" may vary depending on the form of the pharmaceutical composition, the severity of the disease, and the combination with other drugs.

The terms "preventing", "treating" or "ameliorating" as used herein include delaying and stopping the progression of the disease, or eliminating the disease, and do not require 100% inhibition, elimination and reversal. In some embodiments, the inventive traditional Chinese medicine composition or pharmaceutical formulation is capable of preventing the occurrence of RA and/or RA-induced multi-organ complications, or reducing, inhibiting and/or reversing existing RA and/or RA-induced multi-organ complications by, for example, at least about 10%, at least about 30%, at least about 50%, or at least about 80% compared to the levels observed in the absence of the inventive traditional Chinese medicine composition or pharmaceutical formulation.

As used herein, the terms "parts by weight" and "parts by weight" are used interchangeably and the parts by weight may be any fixed weight in milligrams, grams, or kilograms (e.g., 1mg, 1g, 2g, or 1kg, etc.). For example, a composition comprising 1 part by weight of component a and 9 parts by weight of component b may be a composition comprising 1 gram of component a+9 gram of component b, or 10 grams of component a+90 gram of component b, etc. In the composition, the percentage content of a certain component= (the parts by weight of the component/the sum of the parts by weight of all components) ×100%, and therefore, in the composition composed of 1 part by weight of component a and 9 parts by weight of component b, the content of component a is 10%, and the content of component b is 90%.

As used herein, the term "raw medicinal material" refers to a traditional Chinese medicinal material that is an active medicinal material in the composition of the present invention, and the medicinal material may include various forms, for example, the medicinal material includes (but is not limited to) dry medicinal material, fresh medicinal material, raw medicinal material, cooked medicinal material, processed medicinal material, or a combination thereof. In addition, when the composition of the present invention contains a certain crude drug extract, the term also includes the crude drug corresponding to the crude drug extract or the corresponding crude drug used for preparing the extract.

As used herein, the term "feed to liquid ratio" refers to the ratio (w/v) of the mass of the "feed" in the solid state to the volume of the "liquor" as the leach liquor.

Rheumatoid arthritis and complications of multiple organs caused by rheumatoid arthritis

Rheumatoid arthritis (Rheumatoid Arthritis, RA) is an autoimmune disease, and the pathology of RA is mainly synovial lining cell proliferation, massive inflammatory cell infiltration of the interstitium, and the neovascularization, pannus formation, destruction of cartilage and bone tissue, etc.

Rheumatoid arthritis is susceptible to complications, including but not limited to: kidney disease, skin disease, respiratory disease, eye disease, heart disease, liver disease, splenomegaly and anemia.

More than half of RA patients develop serious complications, wherein interstitial lung disease occurs in the lungs, the disease is poorly controlled, diffuse lung fibrosis and alveolus lung can develop, and the patients develop respiratory dysfunction, gas-oxygen exchange disorder, respiratory distress and finally die due to respiratory failure.

The heart may be complicated with cardiomyopathy, coronary vasculitis, arrhythmia and valvular disease, heart failure and ischemic heart disease. The increase in mortality in RA patients is largely due to the occurrence of cardiovascular complications (about 30% -40%).

It has also been found clinically that the incidence of autoimmune liver disease is increased in RA patients, with 50% of RA patients experiencing elevated liver enzymes, and more than 60% of RA patients may have portal chronic low-grade inflammatory infiltrates and small foci of hepatocyte necrosis, with a incidence of primary biliary cirrhosis of up to 10%.

Chronic kidney disease is also common in RA patients with a 37% -57% probability of complications. Wherein mesangial proliferative glomerulonephritis, membranous glomerulonephritis, and immune nephropathy occur at a higher rate in renal complications in RA patients. The occurrence of secondary amyloidosis in severe cases causes renal failure.

Patients with RA also experience symptoms of splenomegaly and anemia, and severe cases even experience spleen rupture, endangering the life of the patient. These complications lead to a 3-fold increase in the risk of mortality in RA patients, with a 10-15 year reduction in the average life span.

As used herein, the terms "multi-organ complication" and "multi-organ complication due to RA" are used interchangeably to refer to damage to organs such as the lung, heart, liver, kidney, spleen, etc. caused by rheumatoid arthritis. The multi-organ complication may be a lesion of a plurality of organs caused by rheumatoid arthritis, or a lesion of a single organ caused by rheumatoid arthritis, wherein the organs include but are not limited to lung, heart, liver, kidney and spleen.

The traditional Chinese medicine composition of the invention

The invention provides a traditional Chinese medicine composition for preventing, treating and/or improving RA and/or multi-organ complications caused by RA.

Study theory support: the traditional Chinese medicine considers that rheumatoid arthritis and visceral complications thereof belong to the categories of five-body arthralgia and five-viscera arthralgia in the theory of arthralgia syndrome, and the yellow emperor internal channel and element question is described as follows: wind-cold-dampness mixed with three qi is also known as arthralgia. "Su Zhi Fang Ji Si" (plain question): the arthralgia and the blockage also cause the bleeding and the qi to be coagulated and unsmooth. "Huangdi's interior channel, su Bian": the five zang organs are all combined, and the patients with long-term illness are also in the combination. "in Nei Jing": the five bodies are all combined with the five zang organs, reflecting that there is a correspondence between the five bodies and the five zang organs. The body arthralgia is not healed for a long time, and the body is infected with exogenous evil again, and the disease is transmitted into the interior, resulting in five viscera arthralgia. Five-body arthralgia can cause five-viscera arthralgia and can also be transmitted and changed mutually. In the course of pathological changes, the human body is deficient in vital qi, and when the human body is invaded by exogenous pathogenic factors, the disease is aggravated by blood stasis, wind-dampness and blockage of channels and collaterals, so that the disease is built in the five zang organs.

The blockage of the channels and collaterals is the root cause of complications of multiple organs caused by RA, and the treatment method for tonifying qi, activating blood, dispelling wind, removing dampness, dredging the arthralgia and relieving pain can achieve better clinical curative effect.

And (3) square solution: the Chinese medicine for treating the arthralgia is prepared from the Chinese medicinal materials Shi Qi teaching the concept of treating injuries of Shanghai university, mainly including qi and blood, and is prepared from radix astragali, chinese angelica and spleen and blood, and is taken as a monarch drug; herba et Gemma Agrimoniae enters lung, liver and spleen channels and nourishes blood, herba Epimedii walks liver and kidney channels and is an essential drug for invigorating vital gate, replenishing vital essence and qi, strengthening tendons and bones, tonifying kidney and strengthening yang, and herba et Gemma Agrimoniae and herba Epimedii are ministerial drugs for replenishing blood and tonifying kidney; both bupleurum and gentiana macrophylla enter liver and gall channels and play roles of dispersing and regulating liver qi, dispelling wind-damp, clearing heat and relieving arthralgia together as adjuvant drugs; rhizoma corydalis is combined with Cyathula root as an guiding drug for activating blood and dredging meridians. The whole formula has the effects of tonifying qi, activating blood, dispelling wind, removing dampness, relieving arthralgia and relieving pain. The recipe is often used clinically for treating deficiency of vital energy, blood stasis and wind-damp, obstruction of channels and collaterals, shoulder pain, arm pain, lumbago and skelalgia, or general pain, with long-term cure.

The invention adopts a compound preparation of the Chinese medicinal composition prepared by the professor Shi Qi of Shanghai university of Chinese medicine and prepared from astragalus, angelica, bupleurum, gentiana macrophylla, corydalis tuber, achyranthes, hairyvein agrimony and epimedium herb extracts to treat the complications of multiple organs caused by RA.

In the invention, a preferable composition of the 'dirty arthralgia formula' is 5-50 g of radix astragali, 3-30 g of Chinese angelica, 3-30 g of radix bupleuri, 3-30 g of gentiana macrophylla, 3-30 g of rhizoma corydalis, 3-30 g of radix cyathulae, 3-30 g of herba epimedii and 3-30 g of hairyvein agrimony.

In the invention, a preferable composition of the 'dirty arthralgia formula' is prepared from 18 g of astragalus root, 12 g of Chinese angelica, 9 g of bupleurum root, 9 g of gentiana macrophylla, 6g of corydalis tuber, 12 g of medicinal cyathula root, 12 g of epimedium herb and 12 g of hairyvein agrimony.

In the invention, a preferable composition of the 'dirty arthralgia formula' is 30 g of astragalus root, 12 g of angelica, 12 g of bupleurum, 12 g of gentiana macrophylla, 9 g of corydalis tuber, 12 g of medicinal cyathula root, 12 g of epimedium herb and 12 g of hairyvein agrimony.

The extractant of the invention and the preparation method thereof

The invention provides an extracting agent of a traditional Chinese medicine composition, wherein the extracting agent is prepared by extracting the traditional Chinese medicine composition with water and/or ethanol.

As used herein, the "extractant of the present invention" refers to an extract in the form of a decoction or an extract obtained by purifying the traditional Chinese medicine composition of the present invention by extraction with ethanol or an aqueous solvent and alcohol precipitation and filtration.

In the present invention, a preferred extractant is a concentrated decoction or extract having a relative density of 1 to 1.4, preferably 1.05 to 1.35 or 1.3 to 1.4.

The invention also provides a preparation method of the extractant,

1) The method comprises the following process steps:

weighing the raw materials according to the weight ratio, soaking, adding 8-14 times of water, decocting for 2-3 times, each time for 1-3 hours, mixing decoctions, filtering, concentrating the filtrate into extract with the relative density of 1.05-1.35 for later use;

adding ethanol into the extract to make the ethanol concentration be 50-80%, standing overnight, concentrating supernatant to relative density of 1.3-1.4;

2) The method comprises the following process steps:

weighing the raw materials according to the weight ratio, soaking, adding 8-10 times of 50-90% ethanol, decocting for 2-3 times, each time for 1-3 hours, mixing decoctions, filtering, concentrating the filtrate into extract with the relative density of 1.05-1.4 for later use;

drying the concentrated extract under reduced pressure, and pulverizing to obtain extract powder;

3) The method comprises the following process steps:

weighing the raw materials according to the weight ratio, soaking, decocting for 2 times, each time for 1-3 hours, adding 50-90% ethanol for 8-10 times in one decoction, adding water for 6-10 times in the other decoction, filtering the decoction, and concentrating to obtain extract with the relative density of 1.3-1.4 for later use;

the active ingredients of the traditional Chinese medicine composition are purified by ethanol or water solvent extraction and alcohol precipitation and filtration, wherein the adopted extraction method is to select the extraction solvent and extraction conditions of alcohol precipitation and filtration according to the physicochemical properties of the active ingredients in the medicine; the adopted purification method adopts ethanol precipitation method to carry out purification treatment on the premise of ensuring curative effect.

Formulations and administration

The present invention provides a formulation including, but not limited to: the pharmaceutical preparation or health product preparation is prepared from one or more medicinal materials or extracts thereof, such as radix astragali, radix Angelicae sinensis, bupleuri radix, radix Gentianae Marcrophyllae, rhizoma corydalis, radix Cyathulae, herba Epimedii, and herba et Gemma Agrimoniae. In the preparation, the content of the raw medicinal materials or the extracts thereof can be 0.1-99.9wt% based on the weight of the preparation. The composition may also contain a pharmaceutically or nutraceutically acceptable carrier.

In the present invention, it should be understood that the extract includes not only a concentrated solution (e.g., an extract) or an active ingredient (e.g., a powder) obtained by extracting a mixed raw material, but also a concentrated solution (e.g., an extract) or a mixture of active ingredients (including a mixture between the concentrated solution and the concentrated solution, between the active ingredient and the active ingredient, or between the concentrated solution and the active ingredient) obtained by extracting a single raw material. The active ingredients of the raw medicinal materials comprise single components or mixed components.

The dosage form of the formulation of the present invention is not particularly limited, and may be any dosage form suitable for mammals. Preferably, the dosage form may comprise a tablet, capsule, granule, pill, mixture, powder, oral liquid, buccal tablet, or aerosol. The preferred composition is a solid formulation from the standpoint of ease of preparation, administration or administration. Oral administration is preferred.

Various conventional carriers and/or auxiliary materials required for preparing different dosage forms can be added into the preparation. The auxiliary materials in the invention are pharmaceutical standard auxiliary materials, including dextrin, lactose, sodium bicarbonate, citric acid, hydroxymethyl starch sodium, micro powder silica gel, starch, soluble starch, polyethylene glycol or magnesium stearate. Can be prepared into any common dosage forms such as tablets, capsules, granules, capsules, pills and powder by adopting a conventional traditional Chinese medicine preparation method.

Examples of pharmaceutically acceptable excipient moieties are cellulose and its derivatives (e.g. methylcellulose, ethylcellulose, hydroxypropylmethyl cellulose, sodium carboxymethylcellulose, etc.), gelatin, talc, solid lubricants (e.g. stearic acid, magnesium stearate), calcium sulphate, vegetable oils (e.g. soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (e.g. propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifying agents (e.g. tween), wetting agents (e.g. sodium lauryl sulphate), buffering agents, chelating agents, thickening agents, pH-adjusting agents, transdermal enhancers, colorants, flavouring agents, stabilizers, antioxidants, preservatives, bacteriostats, pyrogen-free water, etc.

The preparation can avoid temporary decoction before taking the traditional Chinese medicine decoction, is easy to mildew and deteriorate after long-term placement and brings a lot of inconvenience to patients, and according to clinical medication characteristics, the physicochemical properties of main medicinal ingredients in the prescription are preserved, and the preparation is prepared into solid preparations such as granules, tablets, capsules, powder and the like by combining with modern preparation technology, so that the requirements of patients are conveniently and effectively met.

In a preferred form, the formulation corresponds to 5-50 g, preferably 10-20 g, of total medicinal material per gram or ml calculated on the total dry weight of the medicinal material.

In a preferred embodiment, the preparation method of the formulation of the present invention comprises the steps of:

drying the concentrated extract under reduced pressure, and pulverizing to obtain extract powder; adding medicinal auxiliary materials into the extract powder, uniformly mixing, granulating, and pressing into 10 tablets, wherein each tablet is 0.45g; or making into granule, capsule, mixture or powder.

The preparation of the invention can be directly used for preparing medicines for preventing and/or treating RA and/or multi-organ complications caused by RA. The preparation of the present invention may further contain a drug derived from other optional medicinal materials or an extract thereof or other drugs effective for treating and/or preventing RA and/or multi-organ complications caused by RA. Preferably, the formulation of the invention may alleviate RA and/or multiple organ complications caused by RA resulting from antigen stimulation, wherein the antigen comprises lipopolysaccharide, virus, bacterial toxins. More preferably, the formulation of the present invention may alleviate RA and/or RA-induced complications of multiple organs caused by lipopolysaccharide stimulation.

Typically the formulations of the invention are administered at a dose of 0.001-20g/kg (preferably 0.5-1.5 g/kg) of animal body weight per day, preferably at 1-4 divided doses per day, or in a slow release form. For most mammals, the total daily dose is 0.05-600g. Of course, the particular dosage will vary with the mode of administration, the dosage form and the severity of the condition being treated, and these are within the skill of the skilled practitioner. Administration may be by conventional routes including, but not limited to: oral, intramuscular, dermal, or topical administration. Oral administration is preferred.

The present invention also provides a method for preventing and/or treating RA and/or RA-induced multi-organ complications, said method comprising the steps of: the compositions or formulations of the present invention are administered to a subject in need thereof. The subject may be a human or non-human mammal (e.g., dog, pig, cat, monkey, sheep, horse, cow, etc.).

The main advantages of the invention include:

1) The traditional Chinese medicine composition has definite curative effect and high safety;

2) The preparation method of the traditional Chinese medicine composition is simple, low in cost and easy to obtain raw materials;

3) The traditional Chinese medicine composition can reduce inflammatory cell infiltration and pulmonary fibrosis of the lung, improve myocardial hypertrophy and heart fibrosis, reduce hepatic vascular inflammation and hepatic fibrosis, and reduce renal interstitial inflammation, fibrosis and glomerulopathy and splenomegaly;

4) The traditional Chinese medicine composition can simultaneously improve interstitial pneumonia, cardiovascular diseases, autoimmune liver diseases, chronic nephritis and splenomegaly caused by RA and treat multi-organ inflammation caused by RA.

The invention will be further illustrated with reference to specific examples. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. The experimental methods, in which specific conditions are not noted in the following examples, are generally conducted under conventional conditions or under conditions recommended by the manufacturer. Percentages and parts are weight percentages and parts unless otherwise indicated.

The quality of the traditional Chinese medicine materials related by the invention accords with the relevant regulations under a corresponding medicinal material item of 2010 edition of Chinese pharmacopoeia.

Example 1.

5-50 g of raw astragalus, 3-30 g of Chinese angelica, 3-30 g of radix bupleuri, 3-30 g of gentiana macrophylla, 3-30 g of corydalis tuber, 3-30 g of medicinal cyathula root, 3-30 g of epimedium herb and 3-30 g of hairyvein agrimony.

Soaking the raw materials for 1h, decocting for 2 times, each time for 1h, adding 14 times of water for one time and 10 times of water for the other time, mixing decoctions, filtering, and concentrating the filtrate to obtain extract with relative density of 1.05-1.07;

adding ethanol into the extract liquid to make the ethanol concentration be 50-80%, standing overnight, concentrating supernatant until the density is 1.3-1.4;

drying the concentrated extract under reduced pressure, and pulverizing to obtain extract powder. Adding medicinal adjuvants into the extract powder, mixing, granulating, and pressing into 1000 tablets, each tablet being 0.45g, and taking 10 tablets each day. Or making into granule, capsule, mixture or powder according to conventional pharmaceutical method.

Example 2.

18 g of raw astragalus, 12 g of Chinese angelica, 9 g of bupleurum, 9 g of gentiana macrophylla, 6g of corydalis tuber, 12 g of medicinal cyathula root, 12 g of epimedium, and 12 g of hairyvein agrimony.

Each of the crude drugs was prepared as described in example 1 to obtain extract powder. Adding medicinal adjuvants into the extract powder, mixing, granulating, and pressing into 1000 tablets, each tablet being 0.45g, and taking 10 tablets each day. Or making into granule, capsule, mixture or powder according to conventional pharmaceutical method.

Example 3.

30 g of raw astragalus, 12 g of Chinese angelica, 12 g of radix bupleuri, 12 g of gentiana macrophylla, 9 g of rhizoma corydalis, 12 g of medicinal cyathula root, 12 g of herba epimedii and 12 g of hairyvein agrimony.

Example 4.

Weighing the raw materials according to the weight ratio described in the embodiment 1, soaking, adding 8-10 times of 50-90% ethanol, decocting for 2-3 times, each time for 1-3 hours, mixing decoctions, filtering, concentrating the filtrate into extract with relative density of 1.05-1.4 for later use;

drying the concentrated extract under reduced pressure, and pulverizing to obtain extract powder.

Example 5.

Example 6.

30 SPF-grade male DBA/1 mice of 3 month-old cleaning grade were randomly divided into a normal control group, a model+physiological saline group, a model+visceral arthralgia prescription group (also called a treatment group, a tablet prepared in example 1 was randomly selected, and dissolved in water) according to body weight.

Normal control mice were untreated and model mice were daily injected with chicken type ii collagen containing five monoclonal antibodies from the tail vein. Five days later, 25ug Lipopolysaccharide (LPS) was injected intraperitoneally and the detection of joint inflammation peaked at day 15. And (5) finishing the molding.

The treatment group carries out the gastric lavage of the model group with the dirty arthralgia formula of 0.116g/kg every day, and the normal control group and the model+normal saline group carry out the gastric lavage with the same amount of normal saline every day. The lung, heart, liver, kidney, spleen tissues were removed 4 months after the initial administration, respectively, and after formalin fixation and dehydration, HE staining and Masson staining were performed.

Cardiac tissue paraffin sections were also subjected to immunofluorescent staining with Wheat Germ Agglutinin (WGA) specifically staining the myocardial cell membrane to determine myocardial cell hypertrophy based on myocardial cell contours.

Kidney tissue paraffin sections were stained for podocyte (Nephrin) and glomerular mesangial (anti-NG 2 chondroitin sulfate proteoglycan antibodies) immunofluorescence, and kidney lesions were judged for progression based on podocyte and glomerular mesangial proliferation. Meanwhile, the kidney can also be used for PAS staining to observe the change of the basal lamina and the basal lamina.

6.1 in vivo lung HE staining detection results of animal experiments

As shown in fig. 1, the model + saline mice were diffused with mild to moderate inflammatory infiltrates in the lung interstitial, vascular and airway tissues, as compared to the normal control mice. The wall of the middle and small artery is thickened, and the lumen is lined with feather-like epithelial cells.

Compared with the model and normal saline group, the inflammatory area of the model and the visceral pain prescription group is obviously reduced. The research result shows that the viscera obstruction formula can reduce the inflammation degree of lung tissues of interstitial lung diseases.

6.2 in vivo lung Masson staining results of animal experiments

As shown in fig. 1, compared with the normal control mice, the model + normal saline mice showed large areas of collagen fibrous tissue proliferation in the lung bronchi, pulmonary interstitial spaces and perivascular areas.

Compared with the model and normal saline group, the pulmonary fibrosis area of the model and the dirty arthralgia formula group is obviously reduced. The research result suggests that the viscera obstruction formula can reduce the fibrosis degree of lung tissues of interstitial lung diseases.

6.3 in vivo cardiac HE staining results of animal experiments

As shown in the HE staining results of FIG. 2, compared with the normal control mice, the heart tissue of the mice in the model+normal saline group showed hypertrophy of cardiac myocytes and irregular arrangement.

Compared with the model and normal saline group, the myocardial hypertrophy condition of the model and the viscera obstruction formula group is obviously relieved. The research result suggests that the visceral arthralgia formula can relieve myocardial hypertrophy caused by RA.

6.4 results of in vivo cardiac Masson staining in animal experiments

As shown in Masson staining of fig. 2, the heart tissue of mice in the model + saline group was seen as significantly blue collagen fibers compared to normal control mice.

Compared with the model and normal saline group, the heart fibrosis area of the model and the viscera obstruction formula group is obviously reduced. The research result suggests that the visceral pain recipe can significantly reduce the degree of cardiac fibrosis caused by RA.

6.5 in vivo liver HE staining results of animal experiments

As shown in the HE staining results of fig. 3, compared with the normal control mice, the model + saline mice showed inflammatory cell infiltration around the liver blood vessels, and liver sink area inflammatory cell infiltration was accompanied with interfacial inflammation.

Compared with the model and normal saline group, the perivascular inflammatory cell infiltration and the inflammatory cell infiltration of the catchment area of the model and the visceral paralysis group are clearly reduced. The research result suggests that the viscera obstruction formula can relieve inflammatory cell infiltration around liver blood vessels and a manifold area caused by RA.

6.6 in vivo liver Masson staining results of animal experiments

As shown in FIG. 3, the rats in the model +physiological saline group showed large blue-stained collagenous fiber tissue around liver blood vessels, suggesting local group weaving necrosis, compared with the rats in the normal control group. At the same time, the hepatic artery and vein in the collecting tube interval fibroses collagen fiber hyperplasia.

Compared with the model and normal saline group, the fibrosis area in the liver of the model and the viscera obstruction formula group is obviously reduced. The research result suggests that the visceral paralysis prescription can reduce the perivascular fibrosis degree of the liver caused by RA.

6.7 in vivo renal HE staining results from animal experiments

As shown in the HE staining results of fig. 4, compared with the normal control mice, the model + normal saline mice had massive inflammatory cell infiltration of the kidney interstitium, loss of glomerular mesangial structure, proliferation of podocytes and change of cell nucleus morphology, proliferation of capillary membranes, loose renal tubule arrangement, and loss of renal tubular endothelial nuclei.

Compared with the model and the normal saline group, the model and the dirty arthralgia formula group have the advantages that the infiltration condition of kidney interstitial inflammatory cells is definitely reduced, the glomerular mesangial structure is complete, the hyperplasia condition of podocyte and capillary system membranous is reduced, and the arrangement of renal tubules is compact. The research result shows that the visceral paralysis prescription can reduce the kidney interstitial inflammation and the glomerular structure damage.

6.8 in vivo Kidney Masson staining results of animal experiments

As shown in Masson staining of fig. 4, the kidney interstitial substance of mice in model + saline group was seen as large blue-stained collagen fibrous tissue compared to normal control group mice, suggesting local group weaving necrosis.

Compared with the model and normal saline group, the kidney interstitial fibrosis area of the model and the viscera obstruction formula group is obviously reduced. The research result suggests that the visceral paralysis prescription can reduce the degree of interstitial fibrosis of the kidney.

6.9 in vivo spleen HE staining results of animal experiments

As shown in the HE staining results of FIG. 5, the mice in the model + saline group had reduced spleen white marrow, red marrow was engorged, and white marrow was filled with rounded lymphocytes, as compared to the normal control group mice.

Compared with the model and the normal saline group, the model and the viscera arthralgia formula have the advantages that the condition of white marrow reduction and red marrow hyperemia is relieved, and the spleen body shape structure is complete. The research result indicates that the visceral paralysis prescription can relieve splenomegaly of mice.

Discussion of the invention

HE staining results show that the traditional Chinese medicine composition can reduce complications of multiple organs generated after Lipopolysaccharide (LPS) induces CIA model mice. If the lung inflammatory cell infiltration is reduced, the myocardial hypertrophy is improved, the liver vascular inflammation can be reduced, and meanwhile, the renal interstitial inflammation and the splenomegaly can be reduced; masson staining shows that the traditional Chinese medicine composition can relieve fibrosis of heart, lung, liver and kidney of mice induced by Lipopolysaccharide (LPS) in CIA model.

In conclusion, the traditional Chinese medicine composition has excellent effects on lung, heart, liver, kidney and spleen tissues of a rheumatoid arthritis model mouse in the aspects of inhibiting inflammatory cell infiltration, reducing inflammatory areas, reducing tissue structure damage, reducing fibrosis, reducing tissue swelling and the like, and is very suitable for treating rheumatoid arthritis and multi-organ complications caused by the rheumatoid arthritis. It is worth noting that the Chinese medicinal composition of the present invention inhibits multiple organ complications.

All documents mentioned in this application are incorporated by reference as if each were individually incorporated by reference. Further, it will be appreciated that various changes and modifications may be made by those skilled in the art after reading the above teachings, and such equivalents are intended to fall within the scope of the claims appended hereto.

Claims

1. A traditional Chinese medicine composition for treating multi-organ complications caused by rheumatoid arthritis is characterized by being prepared from the following raw materials or extracts thereof:

10-50 parts of raw astragalus mongholicus;

10-30 parts of Chinese angelica;

5-20 parts of bupleurum;

5-20 parts of gentiana macrophylla;

5-20 parts of rhizoma corydalis;

10-30 parts of radix cyathulae;

10-30 parts of herba epimedii;

10-30 parts of hairyvein agrimony.

2. The traditional Chinese medicine composition according to claim 1, wherein in the traditional Chinese medicine composition, the weight ratio of angelica sinensis, radix bupleuri, gentiana macrophylla, radix cyathulae, epimedium and hairyvein agrimony is (0.7-1.2): (0.7-1.2): (0.7-1.2): (0.7-1.2): (0.7-1.2): (0.7-1.2).

3. The traditional Chinese medicine composition according to claim 1, wherein the weight ratio of angelica sinensis, radix bupleuri, gentiana macrophylla, cyathula root, epimedium herb and hairyvein agrimony is 1: (0.75-1): (0.75-1): 1:1:1.

4. the traditional Chinese medicine composition according to claim 1, wherein the traditional Chinese medicine composition is: 18 g of raw astragalus, 12 g of Chinese angelica, 9 g of bupleurum, 9 g of gentiana macrophylla, 6g of corydalis tuber, 12 g of medicinal cyathula root, 12 g of epimedium, and 12 g of hairyvein agrimony.

5. The traditional Chinese medicine composition according to claim 1, wherein the traditional Chinese medicine composition is: 30 g of raw astragalus, 12 g of Chinese angelica, 12 g of radix bupleuri, 12 g of gentiana macrophylla, 9 g of rhizoma corydalis, 12 g of medicinal cyathula root, 12 g of herba epimedii and 12 g of hairyvein agrimony.

6. A traditional Chinese medicine composition extractant for treating multi-organ complications caused by rheumatoid arthritis, which is characterized in that the extractant is prepared by extracting the traditional Chinese medicine composition according to claim 1, wherein the extraction solvent of the extractant is selected from the following groups: water and 50-90% ethanol.

7. The extractant for Chinese medicinal composition according to claim 6, wherein the ratio of the feed liquid of the Chinese medicinal composition to the solvent for extraction is 1:6-20.

8. The extractant for Chinese medicinal composition according to claim 6, wherein the extractant is a decoction or an extract of the Chinese medicinal composition or a concentrated decoction or extract.

9. The extractant for Chinese medicinal composition according to claim 8, wherein the relative density of the extract is 1-1.4.

10. A method for preparing the traditional Chinese medicine composition for treating the multi-organ complications caused by the rheumatoid arthritis according to claim 1 or the traditional Chinese medicine composition extracting agent for treating the multi-organ complications caused by the rheumatoid arthritis according to claim 6, which is characterized in that,

i) The method comprises the following steps:

a1 Providing a crude drug, the crude drug being as defined in claim 1;

c1 Concentrating to obtain extract A;

d1 Mixing the extract A obtained in the step c 1) with ethanol, standing, and concentrating the supernatant twice to obtain an extract B;

alternatively, ii) comprises the steps of:

a2 Providing a crude drug, the crude drug being as defined in claim 1;

b2 Mixing the raw materials with 50-90% ethanol, decocting, and filtering to obtain decoction B;

c2 Concentrating to obtain extract C;

alternatively, iii) comprises the steps of:

a3 Providing a crude drug, the crude drug being as defined in claim 1;

b3 Decocting the raw materials, adding 50-90% ethanol into the first decoction, adding water into the second decoction, and filtering to obtain decoction C;

c3 Concentrating to obtain extract D.

11. A pharmaceutical formulation for treating multiple organ complications caused by rheumatoid arthritis, the formulation comprising:

i) The Chinese medicinal composition of claim 1 or the Chinese medicinal composition extractant of claim 6;

ii) other pharmaceutically acceptable adjuvants, carriers or excipients.

12. The use of the Chinese medicinal composition according to claim 1 or the Chinese medicinal composition extractant according to claim 6 or the pharmaceutical preparation according to claim 11 for preparing a medicament for treating multiple organ complications caused by rheumatoid arthritis.

13. The use according to claim 12, wherein the rheumatoid arthritis is an autoimmune disease resulting from stimulation with an antigen, wherein the antigen comprises lipopolysaccharide, a virus, a bacterial toxin.

14. The use according to claim 12, wherein the multi-organ complication is a complication of an organ selected from the group consisting of rheumatoid arthritis: lung, heart, liver, kidney, spleen, or a combination thereof.

15. The use of claim 12, wherein the multi-organ complication is selected from the group consisting of: interstitial pneumonia, myocardial hypertrophy, autoimmune liver disease, chronic nephritis, splenomegaly, or a combination thereof.

16. The use of claim 12, wherein the treatment of multiple organ complications from rheumatoid arthritis comprises an improvement in one or more indicators selected from the group consisting of:

b) Improving myocardial hypertrophy and/or cardiac fibrosis;

c) Reducing hepatic vascular inflammation and/or hepatic fibrosis;

e) Alleviating splenomegaly.