Pharmaceutical composition for treatment of rheumatism, method for its preparing and using


FIELD: medicine, phytotherapy, pharmaceutical technology, pharmacy.

SUBSTANCE: invention relates to a composition prepared from the following raw medicinal plants: Tripterygium hypoglaucum (Levl.) Hutch, barrenwort (Epimedium), box-thorn matrimony-vine berries (Lycium) and dodder seeds (Cuscuta) taken in the following mass ratio: 1-4; 1-4; 1-4 and 1-4, respectively. Method involves the following procedures: weighing and milling of Tripterygium hypoglaucum (Levl.) Hutch and barrenwort (Epimedium), their separate boiling in water 3 times, wetting box-thorn matrimony-vine berries (Lycium) and dodder seeds (Cuscuta) in warm water at temperature 80-95°C 1-3 times, separate decantation of decocts and liquid, their separate passing through adsorption column, washing out with water up to clearing eluate, elution with 30-99.3% ethyl alcohol and mixing eluate with alcoholic liquid. The total weight of eluate is 1-8-fold more of weight of parent raw medicinal agents. Then method involves recycling each eluent of the raw medicinal agent, concentrating to specific gravity 1.1, drying extracts and their mixing. Pharmaceutical composition is used in treatment of rheumatism, rheumatic arthritis and chronic nephritis. Invention provides realization of indicated designation.

EFFECT: improved preparing method, valuable medicinal properties of pharmaceutical composition.

8 cl, 21 tbl, 13 ex

 

This invention relates to an Antirheumatic remedy, in particular for the pharmaceutical composition, method of its preparation and application.

Rheumatism and rheumatoid arthritis (RA) is an intractable disease, about 18,000,000 patients became disabled due to RA. Investigation of new anti-RA already lasts about a hundred years, aspirin is the first common remedy against RA. Anti-RA mainly can be divided into two types: non-steroidal anti-inflammatory drugs (NSAIDs) and immunosuppressant. NSAIDs include DICLOFENAC SOD or indometacin and adrenocorticotropin. Clinical research shows that NSAIDs is an effective remedy; immunosuppressant and cytotoxic agent include amethopterin, cycloramic, penicillamine, and so forth. In recent years immunological regulation is used in the treatment of rheumatic diseases. All Antirheumatic agents have a relatively serious side effects, up to the present time highly effective with lipotoxicity has not yet been received.

In research and development Antirheumatic means there are three directions. The first direction includes NSAIDs and antagonist of cell factor, such as recombination soluble antagonist of necrotic tumors is of evich factors inhibitor of interleukin-1 and active factor of platelets. The second direction is a new immunosuppressant or immunomodulator, such as cyclosporine A. the Third direction is a complex drug.

In the field of traditional Chinese medicine to treat Bi syndrome (obstruction) (RA) used "Ma-xing-Shi-Gan-Tang (decoction of Chinese ephedra, apricot ordinary, gypsum and licorice Ural)", "Fan-JI-Huang-qi-Tang (decoction Stephanie chetyrehluchevoy and Astragalus)" and "Wu-tou-Tang (decoction of Aconite Sichuan)" medical-pharmaceutical specialist Zhang Zhong-Jing ancient China. "Flowers " torch" is a wild plant in Sichuan province, which during test application in clinical practice in the regions (Sichuan province) proved to a certain extent, the effectiveness for the rheumatics. Unfortunately, found many uncontrolled effects and serious adverse effects on the reproductive system of a man.

There is a long history in Chinese traditional treatment of Bi syndrome, and Chinese traditional treatment of Bi syndrome showed high efficiency, and there are many medications available. In Chinese Dictionaries pharmacy publications 1995 and 2000 edition is mentioned not less than 80 folk remedies and 29 of finished drugs. But there are still a number of problems, mainly: efficiency is the General treatment for severe Bi-syndromes for example rheumatoid arthritis, was not high enough; dosage form of a pharmaceutical preparation does not meet the requirement of modern life, some drugs have a high treatment efficiency, but with serious side effects, such as pharmaceutical drug from Tripterygium wilfordii.

Renowned pharmaceutical composition for the treatment of autoimmune diseases such as rheumatoid arthritis, inflammation and pain, including Tripterygium wilfordii (Patent CN 1098933 from 22.02.95).

The composition has a high treatment efficiency, but has a serious side effects.

Therefore, the development Antirheumatic drugs with high efficiency and lipotoxicity, dosage form which conforms to the use of drugs and modern life, especially Antirheumatic agents, the effectiveness of treatment which is similar to synthetic anti-rheumatic drugs, but with low toxic side effects, is required.

The aim of the present invention is to develop a complex Antirheumatic composition with high efficiency and lipotoxicity and ease of use.

Another purpose of this invention to provide an method of preparation Antirheumatic composition.

The purpose of the claimed invention is achieved by selection with Revich medicinal materials, which prepare the pharmaceutical composition in a variety of dosage forms, including:

Tripterygium hypoglaucum (Levl.) Hutch

and at least one of materials such as

The large mountain woman (Epimedium brevicornum Maxim.),

Berries Wolfberry (Lycium barbarum L.),

The seeds of dodder (Cuscuta chinensis Lam., Cuscuta australis R. Br.)

or their mixture.

The pharmaceutical composition obtained from the above raw medicinal materials taken in different mass ratios.

The claimed composition may consist of Tripterygium hypoglaucum (Levl.) Hutch and at least one of the three medicinal materials.

The optimal mass ratio of these raw medicinal materials according to this invention is:

Tripterygium hypoglaucum (Levl.) Hutch1˜4
The mountain woman large1˜4
Berries Wolfberry1˜4
The seeds of dodder1˜4

Also features a composition consisting of the optimal mass ratio of raw medicinal materials according to the invention:

Tripterygium hypoglaucum (Levl.) Hutch2
The mountain woman large2
Berries Wolfberry1
The seeds of dodder1

It is also proposed composition prepared from two raw medicinal materials in mass ratio:

Tripterygium hypoglaucum (Levl.) Hutch1˜4
The mountain woman large1˜4

The composition is suggested to prepare from these raw medicinal materials in the following weight ratio:

Tripterygium hypoglaucum (Levl.) Hutch2
The mountain woman large2

Proposed composition prepared from raw medicinal materials taken when the mass ratio is:

Tripterygium hypoglaucum (Levl.) Hutch1˜4
The mountain woman large1˜4
Berries Wolfberry1˜4

The optimal mass ratio of the raw medicinal materials for the preparation of the claimed compositions may be:

td align="left"> Berries Wolfberry
Tripterygium hypoglaucum (Levl.) Hutch2
The mountain woman large2
1

The optimal mass ratio of the raw medicinal materials for the preparation of the claimed compositions may be:

Tripterygium hypoglaucum (Levl.) Hutch1˜4
The mountain woman large1˜4
The seeds of dodder1˜4

The optimal mass ratio of the raw medicinal materials for the preparation of the claimed compositions may be:

Tripterygium hypoglaucum (Levl.) Hutch2
The mountain woman large2
The seeds of dodder1

The total content of icariin33H40About15the above pharmaceutical composition is not less than 2.0 mg.

The optimal mass ratio of the raw medicinal materials for the preparation of the claimed compositions may be:

Tripterygium hypoglaucum (Levl.) Hutch1˜4
Berries Wolfberry1˜4 and/or
The seeds of dodder1˜4

The optimal mass ratio of the raw medicinal materials for the prepara the value of the claimed compositions may be:

Tripterygium hypoglaucum (Levl.) Hutch2
Berries Wolfberry1 and/or
The seeds of dodder1

Using the above raw medicinal materials in bearing relation and applying the usual pharmaceutical technology, receive any dosage forms that are acceptable in clinical practice, such as a pill, powder, paste, tablet, capsule, such as a hard capsule and soft capsule, granule, injection and so on.

Another objective of the present invention is achieved by application of the method of preparation of the above-mentioned pharmaceutical composition, comprising the following steps: weighing raw medicinal materials and grinding Triptervgium livpoglaucum (Levl.) Hutch and named the Mountain women large, separate them in boiling water 3 times, soaking these Berries Wolfberry, dodder Seeds with water at a temperature of 80-95°1-3 times, separate transmission received broths and liquids obtained through wide-mouthed resin adsorption column, washing with water of the resin of the column to the clarification of the effluent, the elution 30-99,3% ethanol to the flow of fluid is dark in color, picking up eluent until its color changes from dark to light, the elimination of alcohol W is dasti from the column with water, mixing the eluate with an alcoholic liquid, the total weight of the eluate at 1 to 8 times heavier than the weight of the named raw material medicines, recycling eluent each of medicinal raw material to extract the alcohol concentration to a specific gravity of 1.10, drying of the extracts of each raw medicinal materials separately or in combination, mixing them uniformly or mixing in the proportions obtaining dosage forms acceptable in clinical practice.

Another objective of the present invention is achieved by the fact. what is suggested cooking method named pharmaceutical compositions lies in the fact that Tripterygium hypoglaucum (Levl.) Hutch cut up into small pieces, and extracted three times separately in 13-, 10-, 10-fold waters within one hour. The large mountain woman cut up into segments, extracted separately three times in 15-, 10-, 10-fold waters within one hour. Berries Wolfberry crushed into a coarse material and soaked in 20-fold waters at a temperature of 80-95°C for one hour. The seeds of dodder crumble into coarse powder, soaked in 31-fold waters at a temperature of 80°C for one hour, filtered separately extracts or liquid soaking named four raw medicinal materials then pass through a wide-mouthed resin adsorption Colo is ku, subjected to elution with 70% ethanol to the flow of eluent a dark color, which is collected until its color changes from dark to light, eluent each of the named commodity funds to recycle alcohol extraction, concentrate, dried derived medicinal powders extracts are mixed equally, or in proportion to the receipt of the dosage form, acceptable in clinical practice.

Study the pharmacodynamics of this invented tools (Fansipan-capsule of the above pharmaceutical compositions for the treatment of rheumatism) suggests that the use of Fansipan by perfusion can significantly inhibit primary and secondary injury from arthritis AIDS (AA) the great mouse; significantly inhibit the trailing hypersensitivity reaction (DTH) little mouse ears caused by 2,4-dinitrofluorobenzene (DNFB); significantly inhibit antibody production of hemolysin little mouse and the activity of macrophages, splenocyte 1L-1, 1L-2, 1L-6, TNF little mouse. Fansipan can significantly inhibit the transformation of lymphocytes induced Copom. Fansipan also can significantly inhibit CD4, CD8, in particular noticeable for CD4, and does not significantly affect the ratio of CD4/CD8. The above steps Fansipan have a visible linear dose-response. The minimum efficiency is active dose 12˜ 18 (crude drug)/kg Fansipan also can significantly inhibit the activity of NK cells. However, effective dose Fansipan does not cause atrophy of the thymus, spleen and other immune organs, it also inhibits the activity of macrophage.

Fansipan can significantly inhibit the inflammatory reaction, it can inhibit the increased capillary permeability of the abdominal cavity of a little mouse called acetic acid, to inhibit ear swelling little mouse, caused by Croton oil, to inhibit pleurisy little mouse and aggregation of leukocytes in the capsule CMC great mouse, caused by the gelatin of the eye hondros, but Fansipan has a weak inhibition for swelling of the feet and nails, caused by the gelatin of the eye Chondrus, and camponovo hyperplasia of granulation tissue, a large mouse. In addition, Fansipan can significantly inhibit the reaction heat twist a little mouse called acetic acid.

Experimental example 1.

Impact on arthritis from AIDS (AA)

1.1 Preventive effect on AA big mouse

Choose 72 large SD mice from the same litter, including half female and half male, body weight 180˜220 g, randomly divided into 6 groups, each group of 12 mice. In each cell live 6 mice. Measure the exact narrow measure is through tape maximum perimeters of the foot and ankle of the left and right rear claw as normal values. All mice pour in the stated means of the same volume of the appropriate concentration or inject a solution of gommitnient (Xihuangqi) of the same volume. After 1 hour, spend intradermal injection of complete adjuvant's adjuvant (Freund-FCA) in 0.1 ml of each mouse in the left rear sole. Pour in the tool 1 times a day, continuously for 30 days, every day the same method to measure the perimeters of the foot and ankle of the left and right rear claw large mouse. In this experiment, calculate the degree of swellingchange perimeters, measured before the injection of FCA and after the injection of FCA, the results are presented in table 1.1, 1.2. Upon expiration of the weighed weight of the major organs of animals of different groups, the results are presented in table 1.3, 1.4.

1.1 Impact of Fansipan on the degree of swelling of the ankle joint of the left claw and foot after the injection of FCA big mouse, "
groupdose (g/kg)the degree of swelling3d9d12d14d16d
Id2d
control-0.69±0.170.69±0.120.92±0.180.84±0.411.10±0.301.65±0.682.10±0.55
Fansipan7.50.74±0.120.66±0.0740.83±0.130.77±0.271.11±0.451.34±0.531.91±0.61
Fansipan150.80±0.240.62±0.130.76±0.180.49±0.17*0.73±0.34*1.00±0.48*1.38±0.67*
Fansipan300.75±0.190.67±0.190.87±0.280.63±0.220.73±0.34*0.82±0.43**1.05±0.53**
Triplerygium hypoglalaucum (Levl.)50.72±0.110.68±0.160.91±0.180.66±0.230.88±0.291.03±0.36*1.37±0.33*
Hutch prednisone0.010.64±0.140.64±0.160.50±0.260.46±0.250.72ଐ.46* 0.87±0.46**1.28±0.69*
groupdose (g/kg)the degree of swelling22d24d26d28d
18d20d
control-2.18±0.442.05±0.462.00±0.462.04±0.571.92±0.651.83±0.67
Fansipan7.51.74±0.731.81±0.551.81±0.521.77±0.551.65±0.551.55±0.49
Fansipan151.32±0.59**1.28±0.58**1.34±0.61*1.33±0.67*1.20±0.64*1.08±0.58**
Fansipan300.95±0.50**0.87±0.51**0.95±0.54**0.89±0.59**0.90±0.57**0.86±0.51**
Tripterygium hypoglaucum 51.47±0.43**1.50±0.43**1.49±0.43*1.42±0.53*1.40±0.56*1.32±0.57
(Levl.) Hutch prednisone0.011.18±0.7**61.03±0.67**1.05±0.69*0.90±0.64**0.86±0.65**0.85±0.59**
Compared with control group *P 0.05, **P<0,01 (same in the following)

1.2 Impact Fansipan on the degree of swelling of the ankle joint of the left claw and foot and after injection of FCA for a large mouse onset
groupdose (r/kg)the degree of swelling14d16d18d
2d9d12d
control-0.14±0.050.06±0.100.34±0.360.80±0.521.43±0.671.36±0.61
Fansipan7.50.18±0.060.26±0.360.82±0.521.31±0.641.28±0.71
Fansipan150,15±0.080.02±0.060.13±0.10*0.37±0.31*0.90±0.56*0.79±0.60*
Fansipan300.18±0.090.06±0.060.16±0.08*0.29±0.20**0.49±0.41**0.33±0.29**
Tripterygium hypoglaucum (Levl.) Hulch prednisone50.16±0.070.01±0.070.11±0.100.44±0.19**0.87±0.56*0.84±0.67*
0.010.20±0.060.08±0.080.21±0.160.44±0.430.99±0.630.84±0.74*
groupdose (g/kg)the degree of swelling24d26d28d
20d22d
control-1.28±0.571.38±0.641.35±0.75 1.20±0.781.12±0.63
Fansipan7.51.33±0.711.31±0.731.27±0.731.16±0.731.07±0.65
Fansipan151.74±0.57*1.92±0.61*0.95±0.64*0.88±0.58*1.83±0.55
Fansipan300.27±0.30**0.34±0.31**0.32±0.33**0.31±0.32**0.34±0.32**
Tripterygium hypoglaucum (Levl.) Hulch50.82±0.65*0.89±0.70*0.80±0.67*0.83±0.680.75±0.69
prednisone0.010.82±0.72*0.79±0.74*0.75±0.67**0.68±0.64*0.71±0.67

/tr>
1.3 Impact Fansipan on the change of body weight of large mice that became ill
groupdose (g/kg)the change in body weights (g)
initial body weightthe body weight Ceres month disease AA the increase in body weights
control-228±34231±523
7.5229±34220±46-9
Fansipan15223±40232±349
30224±37256±6032
Tripterygium hypoglaucum (Levl.) Hutch5226±45230±434
prednisone0.01264±55244±31-21
1.4 Impact Fansipan on the scales of immune organs (profilakticheski experiment) great mouse onset:
groupdose (g/kgthe coefficient of organs (g tissue/100 g body weight)
liverspleenthe thymus glandthe adrenal gland
control-3.92±0.650.34±0.100.098±0.0400.027±0.01
Fansipan7.53.73±0.290.31±0.090.078±0.0380.027±0.008
Fansipan153.48±0.320.38±0.100.100±0.0340.023±0.005
Fansipan303.38±0.28*0.44±0.12*0.100±0.0320.022±0.007
Tripterygium hypoglaucum (Levl.) Hutch53.21±0.30**0.36±0.050.052±0.011**0.026±0.009
prednisone0.013.04±0.20**0.32±0.080.050±0.060**0.02±0.004*

1.2 therapeutic effect on AA big mouse

Choose 50 large mice SD male, randomly divide them into 5 groups, treat them the same method, however, begin to pour the tool in mice 12 days after the inflammation caused by the injection of adjuvant FCA, once a day, continuously for 2 weeks, calculate the degree of swellingso: the perimeter on the measurement date minus the perimeter for the current day of the beginning of the infusion means. The results are presented in table 1.5.1.6. scales major organs are presented in table 1.7.

1.5 Therapeutic effect Fansipan on the swelling of the ankle joint of the left claw after the injection of adjuvant FCA for a large mouse onset
groupdose (g/kg)swelling1d2d4d6d
control-1.81±0.271.92±0.192.12±0.222.16±0.27
Fansipan7.51.68±0.501.64±0.541.70±0.571.82±0.61
Fansipan151.44±0.41*1.51±0.36**1.65±0.34**1.74±0.31**
Fansipan301.50±0.561.48±0.41**1.51±0.44**1.59±0.51**
prednisone0.011.78±0.511.70±0.511.63±0.50*1.58±0.50**
groupdose (g/kg)swelling8d10d12d14d
control-1.92±0.321.87±0.341.92±0.391.78±0.44
Fansipan7.51.67±0.681.60±0.711.61±0.771.58±0.71
Fansipan151.46±0.37**1.48±0.30*1.28±0.37**1.22±0.38**
Fansipan301.29±0.58**1.29±0.65**1.26±0.67**1.20±0.68*
prednisone0.011.27±0.46**1.09±0.54**0.94±0.50**0.94±0.42**

0.34±0.32
1.6 Therapeutic effect Fansipan on the swollen ankle right claw after injective adjuvant FCA for a large mouse onset
groupdose (g/kg)swelling2d4d6d8d
control-0.36±0.260.45±0.250.55±0.340.47±0.29
Fansipan7.50.12±0.250.48±0.410.28±0.38
Fansipan150.21±0.180.38±0.270.44±0.330.21±0.33*
Fansipan300.10±0.480.06±0.28**0.11±0.24**0.06±0.27**
prednisone0.010.10±0.13*0.15±0.28*0.11±0.25**-0.08±0.34**
groupdose (g/kg)swelling10d12d14d
control-0.48±0.250.46±0.310.40±0.36
Fansipan7.50.35±0.300.30±0.290.30±0.35
Fansipan150.19±0.45*0.06±0.31**-0.06±0.34**
Fansipan300.02±0.39**0.05±0.38*-0.02±0.41**
prednisone0.01-0.13±0.28**-0.26±0.36**-0.33±0.39**
n=10, compared with the control group, *P<0.05, **P<0.01

1.7 Impact Fansipan on body weight and immune organs(prevention) great mouse onset

groupdose (r/kg)the coefficient of organs(g tissue/100 g body weight)
liverspleenthe thymus glandthe adrenal gland
control-0.35±0.230.35±0.0610.073±0.0140.026±0.0071
Fansipan7.53.21±0.520.33±0.0910.071±0.0260.024±0.0085
Fansipan153.40±0.540.36±0.0140.067±0.0220.023±0.0048
Fansipan302.79±0.430.32±0.0830.069±0.0290,023±0.0072
Tripterygium hypoglaucum (Levl.) Hutch53.92±0.590.35±0.1000.075±0.0340.027±0.0060
prednisone0.013.52±0.350.28±.047* 0.05±0.011*0.02±0.0043*

Tables 1.1, 1.2, 1.3, 1.5, 1.6 show that for a large mouse onset of AA, Fansipan can strongly inhibit the primary damage to the joint after the injection of adjuvant FCA and secondary damage to the joint of the big mouse after injection of the adjuvant FCA. The infusion of funds was effectively at the same time as early inflammation, and 2 weeks after inflammation. This indicates that Fansipan has significant preventive and therapeutic effect on arthritis AA big mouse. Analysis of the impact Fansipan on specific immune swelling of the ankle joint of the hind limbs and nonspecific swelling claw large mouse shows that the action Fansipan on the swollen ankle stronger. This result indicates that the main effect Fansipan is the inhibition of the immune inflammatory response.

The results in tables 1.3, 1.4 and 1.7 shows that during the whole experiment the body weight great mouse onset of AA, has not increased significantly. In the group treated with an effective dose of Fansipan, it was observed an increase in the weight of a large mouse. In the treatment of prednisone and preventive group observed a decrease in the body weight of a large mouse, revealed a marked atrophy of the thymus and adrenal glands. In the group of Tripterygium hypolaucum (Levl.) Hutch also observed atrophy of the thymus, however, in the three groups receiving different doses Fansipan, was not observed noticeable effect on the thymus and adrenal gland.

1.3 Pathological change from AA in the treatment of the stated means of a large mouse

Choose 45 large mice SD and divide them into 6 groups, the body weight of 180±20 g, after the formation of AA through the application's adjuvant adjuvant treatment begins pouring into the stomach Fansipan. Infusion continued for 5 days. Within 1 hour after the last application of funds estimate and calculate the performance of the joint of the big mouse. Joint hindquarters large mice subjected to secondary damage, fixed with formaldehyde. After dyeing is NOT under the microscope were investigated changes in the composition of the synovial membrane and cartilage. The results of the indicators of the composition of different groups large mouse are presented in table 1.8.

1.8 Impact Fansipan on the performance of large joints mouse onset
groupdose (g/kg)the number of mice (thing)indicator replacement
the control group-80**
exemplary group AA-76.2 0.49
Fansipan7.594.86±0.90**
Fansipan1574.71±0.95**
Fansipan3074.56±1.13**
Glucosidorum Tripterygll Totorum0.00674.57±0.79**
Compared with model group, **P<0.01

The degree of swelling of each joint are individually assessed indicators of joint 0-4 points, a set of four points is determined by the index joint. The norm of the estimation of the four limbs and joints as follows: 0 points - normal, 1 point - only red, 2 points - red and slight swelling, 3 points - severe swelling, 4 points - deformation of joints and rigidity.

In the study under the microscope observed hyperplasia of the synovial layer of the joint of the hind limb of a large mouse model group, the increase in collagen fibers, the formation of visible granulomas due to infiltration of lymphocytes and plasma cells; denaturation synovial cells, red cytoplasm and pikes the cell nucleus, the exfoliation of the epithelium of the synovium in some areas; atrophy of cartilage, its surface is rough and uneven, slight hyperplasia of chondrocytes. After treatment with Panipenem different dose gr is PPI may notice relief of inflammation of the synovial tissue of the joint, increasing the formation of collagen fibers; the exfoliation part of synovial cells; hyperplasia of the superficial cells of the cartilage, the surface becomes smooth, the cartilage is in a state of recovery.

The great mouse of the control group may notice hyperplasia of the synovial layer, increase collagen fibers, the formation of visible granulomas due to infiltration of lymphocytes and plasma cells, denaturation of synovial cells, red cytoplasm and pikes the cell nucleus, the exfoliation of the epithelium of the synovium in some areas. After treatment with Panipenem different dose groups celebrated the relief of inflammation of the synovial tissue of the joint, increasing the formation of collagen fibers, exfoliation part of synovial cells, hyperplasia of the superficial cells of the cartilage, the surface becomes smooth, the cartilage is in a state of recovery.

Experimental example 2: effect on the trailing hypersensitivity reaction (DTH) ears, small mouse, called 2, 4-dinitrochlorobenzene (2,4-DNFB)

Choose 50 small mice NIH, including half female and half male, randomly divided into 5 groups. Conduct sensitisation 1% solution of acetone 2.4-DNFB in a dose of 0.025 ml for each mouse on depilation place in the stomach. After three days, increase the sensitization of the same metacone 5 days after sensitization smear right ear little mouse of a 1% solution of edible oil DNFB in a dose of 0.01 ml for each mouse for aggression, after 24 hours kill a small mouse, weighed on a torsion balance the weight of the left and right ear, the difference between them (mg) is the intensity of the DTH reaction of a small mouse. The experiments are carried out in different processes of immunization and receive funds.

2.1 the impact receiving means in the whole process on DTH

The process of immunization and receive funds in the following:

table 2.1.

Influence Pansinin on the trailing hypersensitive reaction little mouse NIH caused
groupdose (g/kg)the deadline means (day)the number of mice (thing)the percentage swelling of the ears (%)the percentage inhibition (%)a value of P
control1034.20±3.77
Fansipan270-51026.24±3.3423.30.01
Fansipan400-51012.99±4.9662.0<0.01
Fansipan600-51010.43&x000B1; 7.5369.5<0.01
dexamethasone0.0030-51013.93±4.4159.3<0.01
control1042.43±5.28
Fansipan40-2-01031.50±10.5225.0<0.01
Fansipan40-2-21030.88±7.9227.20.01
Fansipan40-2-51021.07±4.62*50.30.01
Fansipan405-61032.00±9.3741.70.01
cyclotomic0.05-2-21039.40±10.788.1>0.05
cyclotomic0.05-2-01037.47±6.711 1.7>0.05
control1038.50±4.67
cyclotomic0.1×3once and separately at 0, 2, 4 - day/td> 1023.00±7.6540.30.01
cyclotomic0.25-3d1041.84±7.75-8.7<0.01
Fansipan Cyclotomic600-41027.20±10.2029.4
+ Fansipan0.25+-3,0-41038.07±6.651.1
60
*compared with other groups, P<0.05 or P<0.01

The results in table 2.1 show that Fansipan can significantly inhibit DTH little mouse caused DNFB, the intensity of inhibition markedly depends on the dose, increasing the dose inhibition also increases with the dose of 60.9 g/kg the percent inhibition of DTH is 69.5%.

2.2 Effect of different dates of receiving funds on DTH little mouse

The process of immunization and receive funds and their results are presented in the lower column of table 2.1. The middle graph shows 2 days before sensitization and same-day sensitization, 2 days before sensitization and 2 days after sensitization, 2 days before sensitization through 5 days after sensitization, before and after the aggression, the receiving means can significantly inhibit the reaction, however, the reception means in the whole process before and after sensitization, i.e. two days before sensitization and 5 days after sensitization, shows a stronger inhibition. This indicates that the process and the mechanism of action Fansipan on DTH can be associated with cells involved in the early inhibition of the DTH response, and late effector cells and intermediate cells inhibition of the DTH response. This mechanism differs from cyclopamine. 2 days before sensitization and same-day sensitization or 2 days after sensitization with a small dose of cyclopamine will not affect the DTH reaction.

The bottom graph table 2.1 shows that a single dose of cyclopamine large doses for 3 days before sensitization not only did not inhibit the reaction DTH little mouse, but the response is amplified due to the fact that the function of Th cells relative is activated as a result of the strong inhibition by cyclopamine Ts cells. At this time, if cyclotomic used in conjunction with Panipenem having a noticeable effect of inhibition on DTH, the effect of inhibition Fansipan can be negated. This indicates that Fansipan differs from cyclopamine in the mechanism of action on DTH, because it has high activity in the inhibition of T to ATOC.

Experimental example 3: effects on humoral immunity

3.1 Effect on the antibody production of hemolysin normal little mouse, caused by the immune system of chicken erythrocyte (SCVS)

Choose 190 small mice weighing 18-22 g, half female and half male, randomly divided into 19 groups, each mouse subjected to immunization injection 5% CRBC at a dose of 0.2 ml 7 days after immunization, blood is taken by removal of the eyeball. After dilution of blood samples with saline determine the effect Fansipan on the antibody production of hemolysin little different mouse groups. Infusion Fansipan conducted at different times of immunization. The results are presented in tables 3.1, 3.2, 3.3.

table 3.1

influence Fansipan on the antibody production of hemolysin little mouse
groupdose (g/kg)the deadline meansthe number of mice (thing)the value of hemolysinthe percentage inhibition (%)a value of P
control10169.0±62.0
Fansipan180-710 46.0±15.672.8<0.01
Fansipan270-71035.4±12.079.1<0.01
Fansipan400-71028.2±5.983.3<0.01
Fansipan600-71016.7±3.090.1<0.01
Tripterygium hypoglaucum (Levl.) Hutch13.30-710121.0±88.0**28.4<0.015
cyclotomic0.020-71035.0±12.079.3<0.01
**compared with Panipenem with the same content (40 g/kg) Tripterygium hypoglaucum (Levl.) Hutch P<0.01

table 3.2

Influence Fansipan on the antibody production of hemolysin little mouse
groupdose (g/kg)the deadline meansthe number of mice (thing)the value of hemolysinthe percentage inhibition (%)a value of P
control--1012470± 42.60
Fansipan120-71075.00±53.1039.9<0.05
Fansipan180-71045.60±22.7063.4<0.01
Fansipan270-71029.10±22.1076.8<0.01
Fansipan400-71028.20±5.3077.4<0.01
Tripterygium hypoglaucum (Levl.) Hutch6.00-710143.50±67.90**>0.05
cyclotomic0.020-71027.80±6.6077.9<0.01
**compared with Panipenem the same dose (18 g/kg) P<0.0

table 3.3

influence Fansipan on the antibody production of hemolysin little mouse
groupdose (g/kg)the term means receivingthe number of mice (thing)the value of hemolysinthe percentage inhibition (%)control--10256.0±26.0
Fansipan18-7-710198.0±50.022.7<0.01
Fansipan18-3-710156.0±85.039.1<0.01
Fansipan180-71098.0±35.061.7<0.01
cyclotomic0.020-71025.0±4.090.2<0.01

The results in the above three tables show that Fansipan markedly inhibits antibody production of hemolysin little mouse from different populations, the effect increases with increasing dose. There is a good effect on the dose, the minimum dose inhibition - 12 g/kg, compared with Tripterygium hypoglaucum (Levl.) Hutch, which is the main part of Fansipan has a significantly more powerful effect of the inhibition on the antibody production. The results in table 3.1 show that the intensity of the action Fansipan 2.25 times higher Tripterygium hypoglaucum (Levl.) Hutch. The effect of Tripterygium hypoglaucum (Levl.) Hutch at a dose of 13.5 g/kg weaker than the effect Fansipan content Tripteygium hypoglaucum (Levl.) Hutch in dose 6 g/kg

3.2 Impact on the immunological function of gumor little mouse onset AA

Choose small NIH mice with body weight of 20±2 g, conduct intradermal injection of adjuvant-blockers in the finger of the right hind leg at a dose of 0.05 ml for each mouse. 3 weeks is formed model AA little mouse. After a random division into 6 groups separately pour in different funds within 5 days. Simultaneously with the reception means are subjected to immunization mouse by injection of sheep erythrocytes 10% (CRBC) in a dose of 0.5 ml each, 5 days kill them, take the spleen. After washing fluid Hank's preparing a suspension of lymphocytes, adjusting the concentration of cells to 2×107/ ml. Take 1 ml linfocitos suspension and placed in a test tube by adding 1 ml of 0.2% SRBC and 1 ml of complement 1:30. It is placed in a water bath and incubated for 1 hour at 37°C, centrifuged at 2000 ppm for 5 minutes, take the supernatant and measure 722 spectrophotometer optical density at a wavelength of 415 nm. The value represents the number of PFC.

Another part take blood above sensitised little mouse, sephirot serum agglutination test determine the titer of antibodies, and is represented by Log2 value, the results in table 3.4.

table 3.4

Influence Fansipan on immune is a logical function of gumor little mouse, onset
groupdose (g/kg)the number of mice (thing)PFC (OD)lgM (Log2)
the control group-80.819±0.013#6.875±0.641
exemplary group AA-100.940±0.019**7.700±0.599*
Fansipan580.834±0.012**#6.875±0.641#
Fansipan1080.834±0.012**#6.750±0.886#
Fansipan2080.830±0.014**#6.375±0.518##
Glucosidorum Tripterygll Totorum0.012100.835±0.015**#6.950±0.597#

Compared with control group *P<0.05, **P<0.01; compared with model group, # P<0.05, ## P<0.01.

Table 3.4 shows that the PFC, IgM little mouse onset of AA, is significantly higher than in normal mice. Fansipan can significantly reduce the amount of antibody productive cells (PFC) and the number of education antibodies (IgM) spleen small mouse onset of AA.

Experimental example 4: effect on passive cutaneous anaphylactic reaction (SAR) balsalmic

Take a big mouse and hold intramuscular injection with ovalbumin at a dose of 10 mg/kg, while conducting injection into the abdominal cavity of Bacillus pertussis 2×1010/0.2 ml for immunization. 2 weeks to kill large mouse, take blood and sephirot serum in stock.

Again choose 60 large mice with a body weight of 150˜200 g, half female and half male, randomly divided into 6 groups. Under light ether anesthesia spend intramuscular injection above the serum of antialbumin (degree of dilution is 1:5 and 1:10, in particular, seems d1, d2) 0.1 ml depilation place on the back of the mouse, for each dilution, take 2 injection points. After 48 hours spend aggression intravenous injection of 0.5% physiological solution of Evans blue in a dose of 1 ml containing 1 mg of ovalbumin. After 20 minutes, kill more mice by decapitation, turn the skin of the back of the mouse, on the basis of color depth and square blue spots many people appreciate the category of blue spots on the degree of exudation of the dye. Then cut blue skin into small pieces and soak them in 5 ml of 0.1% solution sernokislotnogo acetone (7:3), after 48 hours spend centrifugation. Take the supernatant and determine the optical density at 590 nm, calculate the degree of reaction PCA great mouse and the percent is inhibition. The results are presented in table 4.

table 4

Influence Fansipan on PCA great mouse
groupdose (g/kg)mark d1d2the ratio of light d1absorption d2
control-5.60±1.782.40±2.460.191±0.1290.096±0.106
Fansipan127.50±2.514.20±2.490.402±0.213*0,192±0.175
Fansipan247.10±2.134.10±1.790,310±0.1770.137±0.099
Fansipan486.00±1.831.70±1.950.121±0.1090.024±0.026*
Tripterygium

hypoglaucum

(Levl.) Hutch
86.11±1.272.56±1.670.223±0.1220.074±0.045
Ketotifen0.12.78±1.64**0.67±1.410.033±0.024**0.027±0.019*
Compared with control group *P<0.05, **P<0.01

Table 4 shows that Fansipan poorly will ingibiruet PCA great mouse only at the high dose compared with the control group, there is a noticeable difference.

Experimental example 5: effect on cell factor

5.1 Effect on TNFα, IL-2 small mouse

Take 60 children ICR mice with a body weight of 18˜22 g, including half female and half male, randomly divided into 6 groups, separately poured into them Fansipan in different doses or other means. The infusion is carried out once a day, continuously for 10 days, within 24 hours after the last dose of means kill them. Under aseptic conditions take the macrophage or splenocyte from the abdominal cavity of a small mouse, wash solution Hank's 2 times, the solution RPIM 1640 without serum 1 times, prepare 2×108/ml cell suspension with the use of a 5% solution of FCS-RPMI 1640, separately add LPS 10 ng/ml or ConAlong/ml carried out the cultivation at a temperature of 37°C in 5% CO248 hours, determine the TNFα or IL-2.

Determination of TNFα:

Take the bar, coated with a monoclonal antibody TNF-α little mouse. In the bar add cultural supernatant dose of 50μl/bore. After 60 minutes of being straps at room temperature add labeled antibody Biotin, at 25°2 hours, then add animalculae streptavidin, after 30 minutes add the substrate. After 30 minutes add dissolve the cliff, determine the value of the OD at a wavelength of 450 nm. Based on the values OD calculate the content of TMF-α (ng/ml) of the standard curve.

Determination of IL-2:

Take in the logarithmic growth period CTLL cells whose growth is dependent on IL-2, cook 1×105/ml cell suspension with the use of a 5% solution of FCS-RPMI 1640. Pour in CTLL cell suspension in a flat-bottomed bar culturing cells with 96 holes 100μl/hole and pour the supernatant for cultivation 100μl/bore, copy each sample 3, simultaneously compare the standard sample rHIL-2 different dilution with nutrient fluid. Under the conditions of 37°WITH 5% CO2cultivate 24 hours, 6 hours before the end of the cultivation centrifuged, allocate the supernatant 110μl/bore add 10 MTTμl/bore, at 37°after 3 hours determine the value of the OD at 570 nm and the value of OD at 630 nm. The final OD value of each sample is in the range OD at 570 nm - OD at 630 nm.

table 5.1

Influence Fansipan on TNF and
groupdose (g/kg)the number of mice (thing)TNF (PG/ml)1L-2 (lU/ml)
control-1087.80±14.6326.30±4.22
121062.14±13.13**16.00±2.89**
Fansipan241058.60±9.63**18.80±2.86**
361054.40±10.88**18.20±2.86**
Tripterygium hypoglaucum (Levl.) Hutch81058.25±10.32**16.00±2.88**
cyclotomic0.021042.20±9.57**10.10±3.00**
*P<0.05, **P<0.01

The results in table 5.1 show that Fansipan significantly inhibits TNFαat a dose of 12 g/kg the effect of inhibition is very noticeable effect increases with increasing dose, but the curve of the dependence of the dose-response flat. For IL-2 Fansipan also has a noticeable effect inhibition, however, the dependence of the dose-response non-obvious.

5.2 Effect on IL-1, IL-6

Take 70 children NIH mice with a body weight of 18˜22 g, including half female and half male, randomly divided into 7 groups. In every pour in Panipenem different dose or other means once a day, continuously for 10 days, within 24 hours after the last pickup is and means, kill them, take the macrophage or splenocyte from the abdominal cavity and determine IL-1 and IL-6.

Determination of IL-1:

Under aseptic conditions take the macrophage or splenocyte from the abdominal cavity, wash solution Hank's 2 times, the solution RPIM 1640 without serum 1 times, prepare 4×106/ml cell suspension with the use of a 5% solution of FCS-RPMI 1640. Take 1 ml of the suspension and poured into centrifuge tubes with round bottom, under the conditions of 37°WITH 5% CO2cultivate 1 hour, allocate unattached cells, add a solution of 5% FCS-RPMI 1640 and LPS (10 ng/ml)under conditions of 37°WITH 5% CO2cultivated for 72 hours, repeatedly freezing and thawing, store at 4°C. Again take a little mouse S under aseptic conditions take thymocytes, cook 1×106/ml cell suspension with the use of a 5% solution of FCS-RPMI 1640.

Take 100μl of cell suspension of thymocytes and 100μl of the supernatant liquid, which was held repeated freezing and melting, and place them in a flat-bottomed bar culturing cells with 96 holes, copy each sample 3, simultaneously compare the standard sample rHIL-1 different degrees of dilution with nutrient liquid, then add ConA 2 ng/hole, under the conditions of 37°WITH 5% CO2cultivated for 72 hours, for 14 hours before the end of cultivation add 3H-TdR 0.1 μ Ci/hole collecting cell mn is kostolny device for collection of cells, determine the value of the cpm.

Determination of IL-6:

Under aseptic conditions take splenetic little mouse, wash solution Hank's on 2 times, the solution RPIM 1640 without serum 1 times, prepare 2×106/ml cell suspension with the use of a 5% solution of FCS-RPMI 1640. Take 1 ml and placed in a centrifuge tube with a round bottom, add ConA (10 ng/ml)under conditions of 37°WITH 5% CO2cultivate 72 hours.

Located in the logarithmic growth period CTLL cells whose growth is dependent on IL-6, enter 1×105/ml cell suspension with the use of a 5% solution of FCS-RPMI 1640.

Put MN cell suspension in a flat-bottomed bar culturing cells with 96 holes 100μl/hole, and pour the supernatant for the cultivation of 25μl/bore, complete solution of 5% FCS-RPMI 1640 200μl/bore, copy each sample 3, simultaneously compare the standard sample rHIL-6 different degrees of dilution with nutrient fluid. Under the conditions of 37°WITH 5% CO2cultivate 72 hours, 6 hours before the end of the cultivation centrifuged, allocate the supernatant 110μl/bore add 10 MTTμl/bore, at 37°after 3 hours, determine the value of the OD at 570 nm and the value of OD at 630 nm. The final value of the OD of each hole is defined in the interval OD at 570 nm - OD 630 nm.

table 5.2

The effect on IL-1 and IL-6 little mouse
groupdose (g/kg)the number of mice (thing)IL-1 (ng/ml)IL-6 (IU/ml)
control-1078.7±7.194.6±6.8
7.51059.3±4.9**64.9±4,8**
Fansipan151053.3±5.7**60.5±4.3**
301054.4±4.8**56.0±4.6**
601047.0±16.6**56.6±6.1**
Tripterygium hypoglaucum (Levl.) Hutch51057.6±4.7**65.7±4.9**
cyclotomic0.02944.5±7.749.6±6.7**

The results in the table show that Fansipan strongly inhibits the formation of macrophages in the production of IL-1 and the formation of splenocytes abdominal cavity during the production of IL-6 little mouse, and the effect increases with increasing dose.

5.3 Effect for the e in the blood plasma NO big mouse starting to get arthritis from AIDS AA

Take 60 large SD mice with a body weight of 160˜200 g, half female and half male, randomly divided into 6 groups. Mice in the control group are subjected to an intradermal injection NS 0.5 ml in the right rear toe. Exemplary group, group Fansipan large, medium and small dose group Glucosidorum Tripterygll Totorum conduct intradermal injection of complete adjuvant's adjuvant (FCA) 0.5 ml of each mouse in the right rear toe. 18 days after the formation of arthritis from AIDS (AA) the great mouse begin to pour into the stomach means 1 times a day, continuously continue 5 days, give distilled water blank control group and model group; individually give Fansipan large, medium and small doses of groups of large, medium and small doses, give tablets Glucosidorum Tripterygll Totorum group positive control group. Within 1 hour after the last treatment take 2 ml of blood from the abdominal aorta, repariruut the blood plasma and keep it at 70°to determine. Determination of NO is carried out in accordance with the explanatory Memorandum jet box NO: take 0.1 ml of blood plasma and add 0.6 ml of reagent C and mixed evenly, then add 0.4 ml of double distilled water and mixed evenly, add Aut 0.1 ml of reagent D and mix evenly incubated on ice for 60 minutes, centrifuged at 12000 rpm for 2 minutes. Take 0.6 ml of the supernatant and add 0.4 ml of double distilled water and 0.1 ml of reagent a And incubated in ice water for about 15 minutes, then add 0.1 ml of reagent B, placed at room temperature for 1 hour, when colorimetrically 545 nm determine the value of OD. Based on the OD values of the samples calculate the content NO NO standard curve, the results are presented in table 5.3.

48
table 5.3

influence Fansipan the blood plasma NO big mouse, Polevaya arthritis from AIDS
groupdose (g/kg)the number of mice (thing)the NO concentrations (μmol/L)(y=Lgx)
the control group-813.55±1.11*1.131±0.032
exemplary group AA-917.56±4.151.235±0.097
Fansipan1270.985±0.087
Fansipan2471.001±0.067
Fansipan71.026±0.062
Glucosidorum Tripterygll Totorum0.006715.25±3.481.173±0.099
Compared with model group, *P<0.05, **P<0.01; compared with group Glucosidorum Tripterygll Totorum

Table 5.3 shows that the level of plasma NO big mouse model group significantly above idle control group. Fansipan can significantly reduce the level of plasma NO big mouse onset of AA, tablet Glucosidorum Tripterygll Totorum can also reduce the level of plasma NO big mouse onset arthritis, but the effect was considerably weaker.

Experimental example 6: Effect on T lymphocytes, NK cells, CD4, CD8 little mouse

6.1 impact on the transformation of lymphocytes in normal small mouse

Take 80 small mice NIH, including half female and half male, randomly divided into 8 groups, separately poured them by different means, once a day, continuously continue 10 days, within 24 hours after the last dose of the means of their kill, in aseptic conditions take splenetic little mouse, wash solution Hank's on 2 times, the solution RPIM 1640 without serum at 1 time cook 2×106/ml cell the suspension with the use of a 5% solution of FCS-RPMI 1640. Pour the cell suspension into a flat-bottomed bar culturing cells with 96 holes 100μl/bore, copy each sample at 3, two holes are injected stimulus (ConA 2 ng/hole) as a transformational holes and another hole not add stimulus as a contrast to the holes. Under the conditions of 37°WITH 5% CO2cultivated for 72 hours, for 14 hours before the end of cultivation add 3H-TdR 0.1μ Ci/hole. Harvested cells are multi-stemmed instrument for collection of cells, determine the value of the cpm, calculate the average value of the copied holes. Directly compare cpm of different groups or indicators of irritation, indicators of irritation calculated by the following formula:

The results are presented in table 6.1.

27.29±7.67
table 6.1

Impact on the transformation of small lymphocytes of the mouse caused by ConA
groupdose (g/kg)the number of mice (thing)cpmfigure irritation
control-1020433±357925.87±3.06
7.51013566±1779**
Fansipan151012708±1692**18.04±3.76
301012809±2575**16.17±4.37
601012090±1706**19.05±3.80
Tripterygium2.51018038±335917.11±2.60
hypoglaucum (Levl.) Hutch51012081±1039**17.58±4.37
cyclotomic0.0299922±1145**13.66±2.28
Compared with control group *P<0.05, **P<0.01

Table 6.1 shows that Fansipan can significantly inhibit the transformation of lymphocytes caused by ConA, and there is a certain dependence of the dose-response.

6.2 Impact on NK cells and CD4CD8normal little mouse

The same experiment with 5.1, within 24 hours after you stop receiving funds prepare 2×108/ml cell suspension a little mouse with the use of a 5% solution of FCS-RPMI 1640. Define CD4CD8and their ratio and NK cells.

Definition CD4and CD8:

Take 50μl suspensions of splenocytes little mouse, add them in the subject is flowed, covered polylysine, and make cell smear. Apply T cells, isolated nylon guns as a positive control. After fixation with acetone block cell smear serum normal little mouse, add biotinamide antibody against CD4CD8at 37°incubated for 2 hours, add animalculae streptavidin at room temperature retain 10 minutes, then add the substrate, after 10 minutes, washed, perform complex color with hematoxylin for 2 minutes. After the gradient dehydration of the alcohol interlock smear gelatin glycerine under a microscope with high magnification believe 200 cells.

The definition of NK cells:

Preparation of cells of the EU: under aseptic conditions take the little mouse splenocytes, wash solution Hank's 2 times, the solution RPIM 1640 without serum 1 times, prepare 2×108/ml cell suspension as the EU with the use of a 5% solution of FCS-RPMI 1640.

preparation of cells TC: take in the logarithmic growth period Yack-1 cells sensitive to NK cells, little mouse, prepare 4×104ml cell suspension as the vehicle.

definition: pour 100μl EU and 100μl TC in flat-bottomed bar culturing cells with 96 holes, copy each sample 3, while comparing EU and T the (EU control:ESμ l+5% FCS RPMI 1640 100μl; control TC:TSμl+5% FCS RPMI 1640 100μl). Under the conditions of 37°WITH 5% CO2cultivated for 24 hours, 6 hours before the end of the cultivation centrifuged, allocate the supernatant 110μl/bore add 10 MTTμl/bore, at 37°after 3 hours determine the value of the OD at 570 nm and the value of OD at 630 nm. The OD value of each hole lies in the interval OD at 570 nm - OD at 630 nm.

The results in table 6.2 show that Fansipan has a certain effect of inhibition on CD4and CD8also there is a dependence of the dose-response, however, the curve dose-response flat. CD4effective dose of 24 g/kg, and for cells CD8only when a large dose of 36 g/kg effect of inhibition noticeable. Accordingly, the ratio CD4/CD8. Fansipan imperceptibly affected. Cycloramic has a strong effect of inhibition on CD4and CD8and the effect on CD8stronger, resulting in a ratio CD4/CD8greatly increased.

For NK cells Fansipan also has a noticeable effect inhibition, however, the dependence of the dose-response non-obvious, but cycloramic has a strong effect of inhibition between the action of cyclopamine at the dose of 20 the g/kg and actions Fansipan at the dose of 12, 24, 36 g/kg there is a very significant difference.

6.3 Impact on the number and function of T lymphocytes little mouse onset AA

Take a little mouse NIH with a weight of 20±2 g, conduct intradermal injection of complete adjuvant Freund''s in her right hind leg at a dose of 0.05 ml, 3 weeks is formed from arthritis AIDS AA. For the negative control group, conduct intradermal injection of saline solution in her right hind leg at a dose of 0.05 ml of the Medium pour in once a day, continuously for 5 days. After 5 days, take separately the peripheral blood of a little mouse different groups and make a blood smear, conduct colouring by lipase. Under oil immersion lens see the percentage of positive cells stained with lipase (percentage of T cells in the peripheral blood). After anesthesia, a small mouse take the spleen and prepare a single cell suspension, washed with a solution of PBS once, allocate the supernatant liquid, add 4 ml of erythrocytes, completely shaken for 2-3 minutes, after complete dissolution of red blood cells centrifuged, allocate the supernatant liquid, washed. After centrifugation allocate the supernatant, settling cell concentration to 1×106/ml, add separately diluted antibody against CD4CD850μl no EV is th pipe, if 4°incubated for 1 hour, after washing, fluorescent wash liquid 2 times add 2 ml of fixing solution. Filter network with 400 cells per tube FCA, analyze through the Flow cytometer, and the results are presented in table 6.3.

Table 6.3 shows that between different groups of ANAE positive cells there is not a noticeable difference, however, cells CD4little mouse onset of AA, increase and cell CD8significantly reduced, the ratio of CD4/CD8noticeable, treatment Panipenem can restore anomaly CD4CD8CD4/CD8to a normal level.

Experimental example 7: Effect on phagocytic function of macrophages peritoneal cavity of a small mouse

Take 50 children NIH mice weighing 18˜22 g, including half female and half male, randomly divided into 5 groups, separately inject medicinal fluid of the same volume of different doses, once a day, continuously for 7 days. Within 1 hour after the last dose of means are injection 10% chicken red blood cells in the abdominal cavity with 0.2 ml each, after 4 hours, kill them, take the peritoneal fluid by drip infusion under a microscope, observe and consider the number of macrophages, faguoqitirute CRBC, and the number of CRBC, the phage is cited by a single macrophage, the results are presented in table 7.

table 7

Influence Fansipan function of favoritemovie CRBC abdominal cavity of a small mouse
groupdose (g/kg)the number of mice (thing)the percentage of favoritemovie (%)indicator favoritemovie
control-1025.75±9.401.28±0.20
Fansipan271033.20±12.771.46±0.36
Fansipan40.51035.20±10.161.21±0.20
Fansipan60.91037.78±20.141.53±0.32
dexamethasone0.005108.33±10.13*1.10±0.18
*P<0.05

The table shows that the dose 27, 40.5 and 60.9 g/kg Fansipan a negligible effect on the phagocytic function of macrophages peritoneal cavity of a small mouse.

Experimental example 8: Effect of increased capillary permeability of the abdominal cavity of a small mouse

Take 90 children NIH mice weighing 18˜22 g, including half of the wives whom whom sex and half male, randomly divided into 9 groups, separately inject medicinal fluid of the same volume of different doses once or once a day continuously for 3 days. Within 1 hour after the last dose of means spend the injection of 0.7% saline solution US into the abdominal cavity, while conducting the injection of 0.5% physiological solution of Evans blue in a dose of 0.1 ml/10 g body weight, 30 minutes to kill the mouse by the loss of the cervical vertebrae, cut the abdominal cavity, wash the abdominal cavity several times with saline 5 ml for each mouse, then download the wash liquid is drained. Then add saline to a volume of 8 ml/mouse, after centrifugation at 3000 pnm take the supernatant, determine the value of the OD at 590 nm, the results are presented in table 8.

table 8

Influence Fansipan to the increased permeability of the capillaries of the abdominal cavity of a little mouse called acetic acid
groupdose (r/kg)reception frequency toolsthe number of mice (thing)the number of effusion paint (OD)a value of P
control--100.29±0.13
Fansipan27qd×1100.26±0.14>0.05
Fansipan40qd×1100.25±0.10>0.05
Fansipan60qd×1100.25±0.09>0.05
control--100.28±0.15
Fansipan27qd×3100.25±0.12>0.05
Fansipan40qd×3100.18±0.10<0.05
Fansipan60qd×3100.15±0.13<0.05
dexamethasone0.15qd×3100.11±0.07<0.01

The results in table 8 show that for increased capillary permeability of the abdominal cavity of a little mouse called acetic acid, after one receiving means Fansipan operates quietly, and after continuous reception means 3 days it has a notable effect of inhibition at her.

Experimental example 9: Effect on exudation of pleurisy and is gregario inflammatory cells little mouse, caused by the gelatin of the eye Chondrus

After the accidental separation of different groups separately carry out the injection of 0.5% physiological solution of Evans blue in 0.1 ml/10 g of body weight in the coccygeal vein, superficial analgesic little mouse ether. Injection make a special needle with 1% solution of gelatin from eye Chondrus in the right thoracic cavity at a dose of 0.03 ml of each mouse. Separately after 4 hours and after 32 hours after inflammation killed by decapitation small mice, cut the abdomen, exposing the aperture. A syringe with a volume of 1 ml twice inject into the abdomen of the rinsing liquid, the sum of which is 2 ml, then collect the wash liquid in the tube. Take 20 ml of the above eluate and add it in 400 ml of leukocyte distributing fluid under a microscope and count leukocytes. Centrifuged residual liquid at 3000 pnm 10 minutes, take the supernatant and determine the optical density at 600 nm, apply the basic solution wash liquid thoracic cavity to check the zero point. The results are presented in table 9.

table 9

Influence Fansipan on the aggregation pleuriticus cells of a small mouse, caused by the gelatin of the eye Chondrus
groupdose (g/kg)the number of cells (2×105)the exudation of the ink (OD)
4 hours32 hour4 hours32 hour
control-46.0±6.916.0±9.60.156±0.0660.109±0.019
Fansipan2726.8±4.5*14.2±8.00.121±0.0620.116±0.031
Fansipan40.510.9±4.0**17.3±4.60.100±0.0480.153±0.032
Fansipan608.0±5.5**6.6±4.7*0.129±0.0660.092±0.051
dexamethasone0.0512.7±10.2**4.4±4.0*0.085±0.0450.063±0.017
*P<0.05, **P<0.01

Table 9 shows that Fansipan can significantly inhibit the aggregation of leukocytes pleurisy little mouse, the action for early aggregation is particularly strong. At the time of 4 h is obtained the regression equation y=44.13-h, r=-0.9625, for later aggregation effect is weak, only at the dose of 20 g/kg there is noticeable effect, and for exudate pleurisy effect is not obvious.

Experimental example 10: effect on the aggregation of leukocytes in the capsule CMC great mouse

Take 64 large SD mice with a weight of 150˜180 g, including half female and half male, randomly divided into 8 groups, separately inject medicinal fluid of the same volume of different doses once or once a day continuously for 3 days. On the day of the experiment, inject 20 ml of 1% CMC solution in the container formed by the prior injection of 20 ml of air into the back of a large mouse in the previous day. After 3.5 h and 7.5 h separately pumped 0.1 ml and poured in a 0.01% solution of brilliant crisologo blue PBS for painting, then under the microscope believe the number of cells in the capsular fluid CMC. The results are presented in table 10

Table 10

Influence Fansipan on the number of cells in the capsule of carboxymethylcellulose great mouse
groupdose (r/kg)quantity (piece)the number of WBC (×107/L)
3.5 hours7.5 hours
control-89.7±4.257.7±17.3
Fansipan27×188.5±3.539.4±16.5
Fansipan40×188.7±7.335.3±23.2
Fansipan60×186.6±3.318.1±8.6**
control-810.97±6.735.6±11.2
Fansipan27×3815.4±9.738.6±15.5
Fansipan40×384.8±3.4**18.4±12.2**
Fansipan60×383.0±2.8**11.0±9.2*
cortisone0.1×3814.2±8.041.7±16.0
control-810.9±3.041.3±6.9
Fansipan18×786.2±3.0*11.4±6.4*
Fansipan27×783.7±1.7**6.4±3.1**
Feng the sun yiping 40×782.5±1.9**5.9±3.9**
cortisone2 mg ×181.5±0.7**3.0±1.0**
Compared with the control group **P<0.01

Table 10 shows that Fansipan can significantly inhibit the aggregation of leukocytes in the capsule CMC great mouse, there is an obvious dependence of the dose-response. With increasing deadline means the act increases, continuous reception means 7 days at a dose of 18 g/kg can be very noticeable to inhibit leukocyte migration, injection of cortisone into the capsule also has a strong effect inhibition.

Experimental example 11: effect on the swelling of the ear little mouse caused cretonne easy oil

Take 60 children NIH mice weighing 18-22 g, half female and half male, randomly divided into 6 groups, separately inject medicinal fluid or mucus gommitnient the same volume of different doses once a day continuously for 3 days. Within 1 hour after the last dose of funds smear evenly two sides of the ear left ear 2% medicine Croton oil 0.02 ml of each mouse. After 4 hours to kill the mouse by the loss of the cervical vertebrae, cut off the left and right ear, weigh weight popolano is about and weight control ear, the difference between them, expressed in mg, is represented as the degree of swelling of the ear, the results are presented in table 11.

Table 11

Influence Fansipan on swollen ear little mouse, caused by Croton oil
groupdose (g/kg)quantity (piece)the degree of ear swelling (mg)the percentage inhibition (%)a value of P
control-1044.38±9.40
Fansipan271039.05±12.3312.00>0.05
Fansipan401036.65±5.8317.64<0.05
Fansipan601034.91±9.7121.34<0.05
dexamethasone0.0031014.13±5.7568.16<0.01

The results in table 11 show that Fansipan significantly inhibited the ear swelling a little mouse called cretonne easy oil, and there is the dependence of the dose-response, but the curve of the dependence of the dose response of the canopy is, at a dose of 13.5 g/kg the effect of inhibition is very noticeable.

Experimental example 12: effect on the reaction heat twist a little mouse called acetic acid

Take 60 small Kunming mice of the genus weighing 18-22 g, half female and half male, randomly divided into 6 groups, separately inject medicinal fluid or mucus gommitnient different doses. Within 1 hour after receiving means inject 0.7% saline solution US into the abdominal cavity 0.2 ml of each mouse. Put a little mouse in a glass pot and watch the latent period of occurrence of the reaction heat twist different mice and the amount of heat twist within 20 minutes, the results are presented in table 12.

Table 12

Influence Fansipan on the amount of heat twist a little mouse called acetic acid
groupdose (g/kg)quantity (piece)the amount of heat twistthe latent period (min)
control-1034.6±14.13.13±0.80
Fansipan271028.2±5.763.82±0.85
Fansipan401031.0±18.43.86±2.00
Fansipan601020.7±12.3*3.95±1.42
Tripterygium hypoglaucum (Levl.) Hutch201025.1±11.93.60±0.93
hydrochloric morphine10 mg/kg100.0±0.00.00±0.00

The results in table 12 show that with a relatively large dose of Fansipan can slow down the appearance of the reaction heat twist a little mouse called acetic acid, and significantly reduce the amount of heat twist a little mouse for 20 minutes. This indicates that Fansipan has some analgesic effect.

Experimental example 13: Effect on the rheological property of the blood of the great mouse onset AA

Choose a large mouse with SD body weight 180±20 g, do intradermal injection of complete adjuvant Freund's finger of the right hind legs 0.05 ml for each mouse, and produced a sample from arthritis AIDS. Do intradermal injection of saline solution into the finger of the right hind legs of 0.05 ml each great mouse negative control group. 3 weeks after the formation of the share samples of mice Obraztsov the group, groups of large, medium and small doses, negative control group and positive control group, give tablets Glucosidurum Tripterygll Totorum positive control group. Give funds infusion once daily and continuously for 5 days. Within 1 hour after the last dose of funds take 3 ml of blood from the abdominal aorta and pour in anticoagulation tube with 1% heparin, determine the viscosity of whole blood conical-plate viscometer type NXE-1 at shear rate 230, 115, 46, 23, 11.5 and 5.75S-1. Determine the viscosity of blood plasma adjustable pressure capillary viscometer type WTP-B; determine the hematocrit and RBC aggregation hematocrite tube by centrifugation, hard indicators of erythrocytes obtained by calculating the above results, the results are presented in table 13.

Table 13 shows that the rheological property of the blood of the great mouse onset of AA, obviously changes, the viscosity of whole blood and blood plasma increases, the hematocrit decreases, the performance of the aggregation of erythrocytes and hard indicators are rising, treatment Panipenem can significantly improve the above indicators rheology.

The above experiments testified about the pharmacological effect of Fansipan, many important farmacologicas what their actions Fansipan has good dependence of dose-response, this indicates that in clinical practice it is possible to adjust the dose to achieve the best treatment.

A study of the effectiveness of treatment Fansipan in clinical practice was conducted in China, Japan and Australia. The results of observations on international standards of diagnosis, treatment and cure of the respective genera disease show that the efficiency of single use capsules Fansipan on RA is about 94%. Significant efficiency is about 60%, capsule Fansipan capable of rapid improvement of the morning ochanine, swelling and other symptoms and sootvetstvujushij indicators of RA, the results are presented in tables 14˜21.

59.38
Table 14

Comparison of the effectiveness of the treatment group with the control group
groupa number of examplesrelief (clinical

the cure)
significantly effectivelyeffectivelynot effectivethe effectiveness of meaningful action (%)efficiency (%)
treatment3251411293.74
control3031012543.3383.33
Table 15

the effect on IgG, IgA, IgM (X±S)
groupa number of examplesIgG before afterIgA before afterW before after
normal3212.45±1.482.37±1.001.58±0.59
treatment3216.92±3.4914.17±1.39**3.65±1.032.39±1.18**1.89±0.881.48±1.01
control3017.03±4.1215.14±2.21**3.45±1.862.32±1.75**2.03±0.951.76±1.28
Compared with the condition before treatment **P<0.0

Table 16

impact on C3, C4 (X±S)
groupthe number of examples (thing)C3C4
toaftertoafter
normal320.62±0.130.14±0.15
treatment321.88±0.720.48±0.12
1.25±0.66**0.26±0.06*
control302.13±0.640.40±0.16
1.56±0.62**0.25±0.07**
Compared with the condition before treatment *P<0.05, **P<0.01
Table 17

impact on ESR, CRP (X±S)
groupquantities of the examples (thing) ESR upafterCRP toafter
normal328.37±5.264.12±1.88
treatment3266.58±9.0130.31±6.53**13.35±6.67
8.86±3.34*
control3073.33±9.0914.21±6.29
35.83±11.61**9.04±3.15**
Compared with the condition before treatment *P<0.05, **P<0.01

Table 18

comparison of hand strength before and after treatment (X±S)
groupmedical group the control group
toaftertoafter
the strength of left hand39.13±20.24 (15)24.00±17.63 (21)
(mmHg)80.47±34.61** (15)55.15±23.27** (21)
power right hand35.85±22.46 (15)22.80±12.32 (21)
85.32±36.32** (15)58.17±20.59** (21)
Compared with the condition before treatment *P<0.05, **P<0.01

Table 19

The effect on swelling and joint pain and morning stiffness (X±S)
subjectmedical groupthe control group
toaftertoafter
swelling and pain of the joint duration of rigor Mortis (min)morning5.79±0.523.14±0.83*5.56±2.153.92#x000B1; 0.26*
50.33±6.4720.24±3.27**48.75±8.3427.50±3.78**
Compared with the condition before treatment *P<0.05, **P<0.01
Table 20

Impact on the transformation of the RF in the negativity
groupa number of examplesnegativity RF
toafterthe percentage conversion to negativity (%)

54.2

44.4
treatment322411

10
control3018

Along with achieving measurable treatment effect, Fansipan can also reduce S1L - 2R, STNF, SIL - 6R serum of patients, the results are presented in table 21.

Table 21

impact on key indicators: SIL - 2R, STNF, SIL - 6R (X±S)
groupthe number of examples (thing)SlL-2R (u/ml)STNF R1 (ng/ml) SIL-6R (ng/ml)
toaftertoaftertoafter
normal32299±681.56±0.4872.05±18.26
(n=32)(n=24)(n=22)
Fansipan15683±1892.87±0.66136.18±28.57
381±157**1.75±0.54**90.15±20.12**
control10765±2032.63±0.72148.21±30.31
412±167**2.38±0.39 (n=8)99.02±26.70**
Compared with the condition before treatment **P<0.01

EXAMPLE 1

TREATMENT of CHRONIC NEPHRITIS RENAL GLOBULES FANSIPAN IN PRACTICE

Studied patients with chronic nephritis 60 people, the main manifestations: albuminar what I bloody urine. Freely divide them into two groups: an experimental group and a comparison group of 30 people. Experimental group: 2 capsules Fansipan each time, three times daily, taken after eating. Comparative group: each time 20 mg launchedtuesday, three times a day, treatment of 8 weeks. Record symptom and change the bodies of patients, urine test: dosage of protein in urine within 24 hours (24h UPQ), white plasma protein (A), liver function (GPT), kidney function (BUN, SCr), the law of blood (WBC). After 4 and 8 weeks of treatment repeat the above test, watch the standard of leadership learning in practice of the new Chinese medicines, appreciate the effect. As a result, in the experimental group of 30 people 18 people have complete remission (60%), 9 people have private remission (30%), 2 people have influence (6,7%), 1 person without the influence (3,3%), the overall efficiency of 96.7%. In the comparative group of 30 people. 15 people have complete remission (50%), 6 people have private remission (20%), 6 people have influence (20%), 2 person without the influence (10%), the overall efficiency of 90%. Compare the significant efficiency of the two groups (complete remission + private remission), P<0.01. In addition, the indicator 24h UPQ (g) in the experimental group faster and more reduced than in the comparison group, before treatment, at the time of treatment 4 weeks and at the moment 8 week the value of 2.63± 0.72, 1.10±0.56 (P<0.05), 0.42=0.28 (P<0.01); values in the comparison group 2.48±0.92, 1.78±0.64, 1.05±0.66 (P<0.01). In the experimental group had 3 people after the reception there was a swelling in the stomach, after eating had taken the medicine, the symptom disappears, in the comparative group, 4 people swelling in the stomach and nausea, have 3 people easy destruction of the liver, the highest value of GPT 68.5 U/L, in 12 patients women with Regulus appeared reduction and disorganization reg.

EXAMPLE 2

TREATMENT OF PATIENTS WITH SLE DISEASE SYSTEMICALLY ACTIVE PERIOD (SLE).

60 patients freely divided into two groups: the experimental group take 2-3 capsules Fansipan each time, three times a day, after receiving food; a comparative group: 50 mg per day of prednisone (every two weeks to reduce by 10 mg when the dosage of 30 mg every two weeks to reduce to 10 mg). Plus injection in the vein of 0.2 g of cyclophosphamide, once in two days. Continuously treated for 3 months. As a result, in the comparative group 2 people were cured, 7 people have considerable efficiency, 14 people have influence, 7 people without influence. In the experimental group practiced cured 3 person 8 person significant efficiency, 13 people have influence, 6 people there is no effect, the patients of the two groups received good effect. Active index SLE ill is x (SLEDAI), precipitation of blood (ESR), the dosage of protein in the urine With324h and the value drops antibodies to ds DNA significantly reduced in the two groups without significant difference. In the comparative group, 5 people white cells are reduced, the 2 people thromobocytopenia, 5 patients nausea, 2 man hair loss, 2 people increase liver function. In the experimental group, receiving Fansipan, small and slight discomfort, only 2 people stomach pain and bloating in the upper part of the stomach, one slight drop in white cells.

Example 3

Woman, 40 years old, abdominal pain, diarrhea with blood, low temperature, weight reduction, point in the abdomen. When checking roentgenometer using agent barium, interior mirror and explore the causes of the disease, identified the disease clone 6 years old.

After treatment with hormones and depressors of the immunity provision of the disease is not stable, sometimes pain in the abdomen, 3-4 times of diarrhea per day, high fever (37.8° (C), the body weight is 46 kg, significant anemia. After taking Fansipan 3-4 capsules once and 3 times a day, after 4 weeks the position of the disease stabilised, after 3 months of treatment abdominal pain and diarrhea disappear, body temperature normal, anemia is reduced, increasing the weight (52 kg). Active index the disease clone (CDA1) is considered cured.

Listed below are Conques is to maintain examples of pharmaceutical compositions according to the invention, which will provide the above effects.

Example 1

The mountain woman large-2222 g

Tripterygium hypoglaucum (Levl.) Hutch 2222 g

Berries Wolfberry 1111 g

The seeds of dodder 1111 g

Crumble Tripterygium hypoglaucum (Levl.) Hutch into small pieces, individually extracted three times in 13-, 10-, 10-fold waters, each time for 1 hour; cut a large mountain woman to pieces, individually extracted three times in 15-, 10-, 10-fold waters, each time for 1 h; crushed Berries Wolfberry in the rough stuff, soaked in 20-fold the warm waters of the 80°1 h; crumble the Seeds of dodder in coarse powder, soaked in 31-fold the warm waters of the 80°1 h; filtered separately broths and liquids soaking four raw medicinal materials, then separately passed through wide-mouthed resin adsorption column and elute 70% ethanol to the flow of eluent a dark color, which is collected until its color changes to a bright color, eluent subjected to recycling to the selection of alcohol. The extract is concentrated and dried.

In the resulting medicated powders extracts add medicinal starch to 200 grams, evenly mixed, then filled into 1000 capsules. Each capsule, manufactured by the claimed method, contains 0.2 g of a mixture of medicinal extracts, and the content of icariin the 33H40O15each capsule is less than 2.0 mg no Pets. The usual dose by mouth is 3 capsules 3 times a day.

Example 2

Tripterygium hypoglaucum (Levl.) Hutch 2000

The mountain woman large 2000

Crumble Tripterygium hypoglaucum (Levl.) Hutch into small pieces, individually extracted three times in 13-, 10-, 10-fold waters, each time for 1 hour; cut a large mountain woman to pieces, individually extracted three times in 15-, 10-, 10-fold waters, each time for 1 h, filtered separately extracts of two medicinal raw materials. Decoctions passed separately through a wide-mouthed resin adsorption column and elute 70% ethanol to the flow of eluent a dark color, which is collected to switch its color to light, eluent subjected to recycling to the selection of alcohol. The extract is concentrated and dried.

Received medicated powders extracts combined and uniformly mixed with medicinal starch, filled into 1000 capsules. Each capsule prepared by the method according to the invention contains 0.2 g of a mixture of medicinal extracts, and the content of icariin33H40O15each capsule is less than 2.0 mg no Pets. The usual dose by mouth is 3 capsules 3 times a day.

Example 3

Tripterygium hypoglaucum (Levl.) Hutch 2000

The mountain woman who rubecula 2000

Berries Wolfberry 1000 g

Crumble Tripterygium bypoglaucum (Levl.) Hutch into small pieces, individually extracted three times in 13-, 10-, 10-fold waters, each time lasts 1 hour; cut a large mountain woman to pieces, individually extracted three times in 15-, 10-, 10-fold waters, each time for 1 hour; crushed Berries Wolfberry in the rough stuff, soaked in 20-fold the warm waters of the 80°1 hour; filter separately broths and soaked fluid three raw medicinal materials, then passed separately through a wide-mouthed resin the adsorption column, elute 70% alcohol before escaping eluent dark color, which is collected to switch its color to light, eluent subjected to recycling to the selection of alcohol. The extract is concentrated and dried.

Received medicated powders extracts combined and uniformly mixed with medicinal starch, filled into 1000 capsules.

Each capsule prepared by the method according to the invention contains 0.2 g of a mixture of medicinal extracts, and the content of icariin33H40O15each capsule is less than 2.0 mg no Pets. The usual dose by mouth is 3 capsules 3 times a day

Example 4

Tripterygium hypoglaucum (Levl.) Hutch 2000

The mountain woman large 2000

The seeds of dodder 1000 g

Crumble Tripterygium hypoglaucum (Levl.) Hutch at IU is the cue pieces separately extracted three times in 13-, 10-, 10-fold waters, each time lasts for 1 hour; cut a large mountain woman to pieces, individually extracted three times in 15-, 10-, 10-fold waters, each time lasts 1 h; crumble dodder Seeds to a coarse powder, heat soaked 31-fold waters 80 for 1 hour, filtered separately fluid otvorenie and soaking three medicinal materials, then are individually large resin adsorption column, then elute 70% alcohol until the eluent flow dark color, which is collected to switch its color to light, eluent subjected to recycling to the selection of alcohol. The extract is concentrated and dried.

Received four of the medicinal powder extracts combined and uniformly mixed with medicinal starch, filled into 1000 capsules.

Each capsule prepared by the method according to the invention contains 0.2 g of a mixture of medicinal extracts, and the content of icariin33H40About15each capsule is less than 2.0 mg no Pets. The usual dose by mouth is 3 capsules 3 times a day

Example 5

Tripterygium hypoglaucum (Levl.) Hutch 2000

The seeds of dodder 1000

Crumble Tripterygium hypoglaucum (Levl.) Hutch into small pieces, individually extracted three times in 13-, 10-, 10-fold waters, each time 1 hour Crumble dodder Seeds to a coarse powder, soaked in 31-fold the warm waters of the 80°1 hour; filter separately broth and soaked liquid two medicinal materials, then passed separately through a wide-mouthed resin adsorption column, then elute with 70% alcohol before escaping eluent dark color, which is collected to switch its color to light, eluent subjected to recycling to the selection of alcohol. The extract is concentrated and dried.

Received two of the medicinal powder extracts combined and uniformly mixed with medicinal starch, filled into 1000 capsules.

Daily dosage capsules prepared in the described manner according to the invention, is equal to the number of medicinal raw materials - 30 g/day.

Example 6

Tripterygium hypoglaucum (Levl.) Hutch 2000

Berries Wolfberry 1000 g

Crumble Tripterygium hypoglaucum (Levl.) Hutch into small pieces, individually extracted three times in 13-, 10-, 10-fold waters, each time for 1 hour; crushed Berries Wolfberry in the rough stuff, soaked in 20-fold the warm waters of the 80°1 hour; filter separately broth and soaked liquid medicinal materials, then passed separately through a wide-mouthed resin adsorption column, then elute with 70% alcohol before escaping eluent dark color, which is collected to switch its color to light, eluent question recircul the application to highlight alcohol. The extract is concentrated and dried.

Received medicated powders extracts combined and uniformly mixed with medicinal starch, filled into 1000 capsules.

Daily dosage capsules made by this invention, is equal to the number of medicinal raw materials - 30 g/day.

1. Pharmaceutical composition for the treatment of rheumatism, cooked from raw medicinal material Tripterygium hypoglaucum (Levl.) Hutch and at least one of the raw medicinal materials selected from the group: a large mountain woman (Epimedium brevicomum Maxim.), Berries Wolfberry (Lycium barbarum L.), Seeds of dodder (Cuscuta chinensis Lam., Cuscuta australis R.Br.), or mixtures thereof, taken in the following mass proportions:

Tripterygium hypoglaucum (Levl.) Hutch1-4
The large mountain woman (Epimedium brevicomum Maxim.)1-4
Berries Wolfberry (Lycium barbarum L.)1-4
The seeds of dodder (Cuscuta chinensis Lam.,
Cuscuta australis R.Br.)1-4

2. The pharmaceutical composition according to claim 1, characterized in that the prepared from these raw medicinal materials taken in the following mass proportions:

Tripterygium hypoglaucum (Levl.) Huch 2
The large mountain woman (Epimedium brevicomum Maxim.)2
Berries Wolfberry (Lycium barbarum L.)1
The seeds of dodder (Cuscuta chinensis Lam.,
Cuscuta australis R.Br.)1

3. The method of preparation of the pharmaceutical composition according to any one of claims 1 and 2, comprising the following steps: weighing raw medicinal materials and grinding Tripterygium hypoglaucum (Levl.) Hutch and named the Mountain women large, separate them in boiling water 3 times, soaking these Berries Wolfberry, dodder Seeds with water at a temperature of 80-95°1-3 times, separate transmission received broths and liquids obtained through wide-mouthed resin adsorption column, washing with water of the resin of the column to the clarification of the effluent, the elution 30-99,3% ethanol to the flow of fluid is dark in color, picking up eluent until its color changes from dark to light, the elimination of alcoholic liquid from the column with water, mixing the eluate with an alcoholic liquid, the total weight of the eluate in 1-8 times heavier than the weight of the named raw material medicines, recycling eluent each of medicinal raw material to extract the alcohol concentration to a specific gravity of 1.10, drying of the extracts of each sirenakersten materials individually or in combination, mixing them evenly or mix in proportion to the receipt of the dosage form, acceptable in clinical practice.

4. The method of preparation of the pharmaceutical composition according to any one of claims 1 and 2 characterized by the fact that Tripterygium hypoglaucum (Levl.) Hutch cut up into small pieces, and extracted three times separately in 13-, 10-, 10-fold waters within one hour, the large mountain woman cut up into segments, extracted separately three times in 15-, 10-, 10-fold waters within one hour, Berries Wolfberry crushed into a coarse material and soaked in 20-fold waters at a temperature of 80-95°C for one hour, the Seeds of dodder crumble into coarse powder, soaked in 31-fold waters at a temperature of 80°in for one hour, filtered separately extracts or liquid soaking named four raw medicinal materials then pass through a wide-mouthed resin adsorption column is subjected to elution with 70% ethanol to the flow of eluent a dark color, which is collected until its color changes from dark to light, eluent of these commodity funds to recycle alcohol extraction, concentrate, dried derived medicinal powders extracts are mixed equally, or in proportion to the receipt of the dosage form, acceptable in clinical practice.

5. How is prigotovleniya pharmaceutical composition according to claim 3, characterized in that get any dosage forms that are acceptable in clinical practice, such as hard capsule, soft capsule, tablet, granule, injection.

6. The method of preparation of the pharmaceutical composition according to claim 4, characterized in that get any dosage forms that are acceptable in clinical practice, such as hard capsule, soft capsule, tablet, granule, injection.

7. The use of the pharmaceutical composition according to any one of claims 1 and 2 to prepare a treatment for rheumatism or rheumatoid arthritis.

8. The use of the pharmaceutical composition according to any one of claims 1 and 2 to prepare funds for the treatment of chronic nephritis.



 

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2 cl, 8 ex

FIELD: organic chemistry, chemical technology.

SUBSTANCE: invention relates to a method for separation of high-molecular lactone-containing compound. Invention describes a method for separation of lactone-containing compounds wherein mixture of lactone-containing compound showing the main chemical structure as 1,14-dihydroxy-12-[2-(4-hydroxycyclohexyl)-1-methylvinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-azatricyclo[22.3.1.04,9]octacoz-18-ene-2,3,10,16-tetraone and at least one among (C2-C6)-alkenyl group and (C1-C6)-alkoxy-group as a side chain and similar compounds is subjected for one or both the following stages in any order: stage (A) of the mixture adsorption on non-ionic adsorbing resin and elution with an aqueous solvent containing silver ions; and stage (B) of the mixture absorption on basic aluminum oxide and elution with organic solvent for separation of each compound. Invention provides the development of effective method for separation of high-molecular compounds being without their chemical structure.

EFFECT: improved method for separation.

14 cl, 2 dwg, 2 tbl, 3 ex

The invention relates to the field inframetrics analysis, to devices for reducing the background signal with conductivity determining ions by ion chromatography

The invention relates to the field inframetrics analysis, and more specifically to the field of devices for preparation of eluents for ion chromatography

FIELD: medicine, pharmacology.

SUBSTANCE: invention relates to a medicinal agent used in treatment of warts. Proposed agent contains an active compound chosen from isopropyl laurate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, ethyl myristate, propyl myristate, butyl myristate and/or ethyl oleate and at least a mixture containing (-)-epicatechol, (-)-epicatechol gallate, (-)-epigallocatechol, (-)-epigallocatechol gallate, (+)-gallocatechol and (-)-gallocatechol gallate possessing the enhanced effectiveness.

EFFECT: valuable medicinal properties of drug.

7 cl, 2 ex

FIELD: method and composition for oral cavity care.

SUBSTANCE: claimed form contains a) 1-40 % of retaining agent, Selected from group containing water soluble hydrophilic resins, water soluble hydrophilic polymers or mixtures thereof, wherein retaining agent is capable of hydration under water or saliva action to produce retention coefficient of 1-4; and b) 60-90 % of local carrier. Moreover composition contains 65 wt.% of water-insoluble particles. Further disclosed is dental paste composition containing a) 30-65 % of water-insoluble particles as retaining agent, wherein solubility of retaining agent in water is less then 1 g/30 g at 25°C; b) 0.01-40 % of active agent for oral cavity care; c) 0.1-25 % of buffer agent. Claimed composition has retention coefficient of 1-4.

EFFECT: improved compositions for oral cavity care.

9 cl, 3 dwg, 5 ex

FIELD: method and composition for oral cavity care.

SUBSTANCE: claimed form contains a) 1-40 % of retaining agent, Selected from group containing water soluble hydrophilic resins, water soluble hydrophilic polymers or mixtures thereof, wherein retaining agent is capable of hydration under water or saliva action to produce retention coefficient of 1-4; and b) 60-90 % of local carrier. Moreover composition contains 65 wt.% of water-insoluble particles. Further disclosed is dental paste composition containing a) 30-65 % of water-insoluble particles as retaining agent, wherein solubility of retaining agent in water is less then 1 g/30 g at 25°C; b) 0.01-40 % of active agent for oral cavity care; c) 0.1-25 % of buffer agent. Claimed composition has retention coefficient of 1-4.

EFFECT: improved compositions for oral cavity care.

9 cl, 3 dwg, 5 ex

FIELD: method and composition for oral cavity care.

SUBSTANCE: claimed form contains a) 1-40 % of retaining agent, Selected from group containing water soluble hydrophilic resins, water soluble hydrophilic polymers or mixtures thereof, wherein retaining agent is capable of hydration under water or saliva action to produce retention coefficient of 1-4; and b) 60-90 % of local carrier. Moreover composition contains 65 wt.% of water-insoluble particles. Further disclosed is dental paste composition containing a) 30-65 % of water-insoluble particles as retaining agent, wherein solubility of retaining agent in water is less then 1 g/30 g at 25°C; b) 0.01-40 % of active agent for oral cavity care; c) 0.1-25 % of buffer agent. Claimed composition has retention coefficient of 1-4.

EFFECT: improved compositions for oral cavity care.

9 cl, 3 dwg, 5 ex

Massage agent // 2302854

FIELD: medicine cosmetic, in particular agent for treatment of cripple infant suffering from infantile cerebral paralysis.

SUBSTANCE: claimed agent contains cameline oil, dogrose oil, and mint oil. Said agent is uniformly distributed on skin, has good absorption, provides good sliding effect, increases blood circulation, relives muscle stress.

EFFECT: agent with increased feeding, anti-inflammation and immunostimulating action.

1 ex

FIELD: bioengineering.

SUBSTANCE: method involves producing acids polysaccharide fucoidan from laminaria by crushing raw material by treating it with 0.5-1.0% food acid solution during 4-5 h and subjecting acid extract to ultrafiltration on membrane of 100-300 kDa with following end product drying by applying lipophilic or spraying or vacuum drying method.

EFFECT: high purity of end product.

FIELD: synthesis of biologically active compounds.

SUBSTANCE: invention provides novel N6-substituted adenine-based heterocyclic compounds depicted by general formula I: , for which meanings of radicals are presented in description, and pharmaceutically acceptable salts thereof manifesting anticancer, mitotic, immunosuppressive, and antiaging activities for vegetable, animal, and human cells, and methods for preparation thereof. Included are also pharmaceutical compositions, cosmetic preparations, and growth regulators, which contain indicated derivatives as active components. Application of indicated derivatives for preparing therapeutical preparations, and cosmetic preparations are also described.

EFFECT: expanded synthetic possibilities in adenine series and increased choice of various biologically active agents.

10 cl, 10 dwg, 9 tbl, 14 ex

FIELD: medicine, pharmacology.

SUBSTANCE: the present innovation deals with creating preparations of psychotherapeutic action. It has been suggested 5 variants of aromatic compositions for normalizing psychoemotional state: for normalizing psychoemotional state in persons with highly unsteady type of the higher nervous activity (HNA), emotionally active; for normalizing psychoemotional state in persons with highly unsteady type of HNA; for normalizing psychoemotional state in persons revealing melancholic, weak type of HNA; for normalizing psychoemotional state in persons with highly steady type of HNA; for normalizing psychoemotional state in persons with weak unsteady type of HNA. The preparations mentioned provide purposeful impact upon affected psychoemotional sphere in persons with different constitutional peculiarities of HNA.

EFFECT: higher efficiency.

5 cl, 15 ex

FIELD: medicine, pharmacology.

SUBSTANCE: the present innovation deals with creating preparations of psychotherapeutic action. It has been suggested 5 variants of aromatic compositions for normalizing psychoemotional state: for normalizing psychoemotional state in persons with highly unsteady type of the higher nervous activity (HNA), emotionally active; for normalizing psychoemotional state in persons with highly unsteady type of HNA; for normalizing psychoemotional state in persons revealing melancholic, weak type of HNA; for normalizing psychoemotional state in persons with highly steady type of HNA; for normalizing psychoemotional state in persons with weak unsteady type of HNA. The preparations mentioned provide purposeful impact upon affected psychoemotional sphere in persons with different constitutional peculiarities of HNA.

EFFECT: higher efficiency.

5 cl, 15 ex

FIELD: medicine, pharmacology.

SUBSTANCE: the present innovation deals with creating preparations of psychotherapeutic action. It has been suggested 5 variants of aromatic compositions for normalizing psychoemotional state: for normalizing psychoemotional state in persons with highly unsteady type of the higher nervous activity (HNA), emotionally active; for normalizing psychoemotional state in persons with highly unsteady type of HNA; for normalizing psychoemotional state in persons revealing melancholic, weak type of HNA; for normalizing psychoemotional state in persons with highly steady type of HNA; for normalizing psychoemotional state in persons with weak unsteady type of HNA. The preparations mentioned provide purposeful impact upon affected psychoemotional sphere in persons with different constitutional peculiarities of HNA.

EFFECT: higher efficiency.

5 cl, 15 ex

FIELD: medicine, pharmacology.

SUBSTANCE: invention relates to a medicinal agent used in treatment of warts. Proposed agent contains an active compound chosen from isopropyl laurate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, ethyl myristate, propyl myristate, butyl myristate and/or ethyl oleate and at least a mixture containing (-)-epicatechol, (-)-epicatechol gallate, (-)-epigallocatechol, (-)-epigallocatechol gallate, (+)-gallocatechol and (-)-gallocatechol gallate possessing the enhanced effectiveness.

EFFECT: valuable medicinal properties of drug.

7 cl, 2 ex

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