WO2006063515A1

WO2006063515A1 - Use of radix sanguisorbae and its extract for preparing medicament to increase rbc and hemoglobin

Info

Publication number: WO2006063515A1
Application number: PCT/CN2005/002176
Authority: WO
Inventors: Bogang Li; Wenjun Zou; Zhongrong Liu; Jianming Wu
Original assignee: Chengdu Di'ao Pharmaceutical Group Co., Ltd.
Priority date: 2004-12-14
Filing date: 2005-12-13
Publication date: 2006-06-22
Also published as: CN1788758A

Abstract

RADIX SANGUISORBAE, its total saponin extract and ursane-type triterpenide saponin of formula (I) are useful for preparing medicament to increase the contents of RBC and hemoglobin. They may be produced into preparation such as oral preparation or injection, with pharmaceutically accepted carriers or adjunctive ingredients. The preparation can increase the contents of RBC and hemoglobin significantly and can treat or prevent all kinds of anaemia, especially for anaemia caused by malfunction or exhaustion of bone marrow.

Description

Application of Chinese medicine mantle and its extract in the preparation of ascending red blood cells and hemoglobin drugs

Technical field

The invention relates to a new use of a traditional Chinese medicine mantle and an extract thereof, in particular to a medicine for preparing an elevated red blood cell and hemoglobin, and belongs to the field of traditional Chinese medicine.

Background technique

Under normal conditions, the generation and decay of red blood cells maintain a dynamic balance, and the values of red blood cells, hemoglobin and hematocrit in a unit volume are kept stable. Reduced or destroyed erythropoiesis, or both, can cause anemia. The anemia caused by the decrease of erythropoiesis mainly includes: 1) Insufficient or utilization of hematopoietic raw materials, such as anemia caused by abnormal iron metabolism, anemia caused by abnormal folic acid or B ₁₂ metabolism; 2) hypofunction or failure of bone marrow hematopoietic function, such as regeneration barrier brick Anemia; 3) Secondary anemia caused by various diseases. Anemia caused by excessive destruction of red blood cells may include red blood cell intrinsic factors such as erythrocyte membrane defects or red blood cell external factors such as immune hemolysis. In addition, excessive blood loss can also cause blood loss anemia. It can be said that anemia is a multiple disease of various subjects such as internal medicine, surgery, pediatrics, obstetrics and gynecology. The treatment of anemia mainly includes drugs, blood transfusion, spleen resection, hematopoietic stem cell transplantation, gene therapy, etc. Generally, one or more treatments are selected according to different etiology or pathogenesis, and drug treatment is still the most important for treating anemia. means. There are many therapeutic drugs for anemia caused by insufficient hematopoietic raw materials or utilization disorders, such as iron-containing preparations, but relatively few drugs for anemia caused by low blood marrow hematopoietic function or failure, such as the treatment of aplastic anemia mainly using males. Hormone or glucocorticoid therapy, such as testosterone propionate and dexamethasone, but these drugs have more side effects, such as testosterone propionate leading to male or prostatic hyperplasia, dexamethasone causing osteoporosis, etc. New biological agents such as erythropoietin (EP0) have a significant clinical effect, but are mainly limited to the treatment of renal anemia, and most of these drugs are injectable and inconvenient to use.

The mantle is the dried root of the Sanguisorba officinalis L. or the long-leaved mantle Sanguisorba officinalis L. Var Longifolia (Bert) Ytl et Li. ("Modern Research and Application of Traditional Chinese Medicine"

(Volume II), Zheng Huzhan, editor, Beijing, Xueyuan Press, 1997, 10), is a traditional Chinese medicine for Chinese medicine, cold, bitter, phlegm, with cooling blood to stop bleeding, detoxification and sore, often used for vomiting blood , hemoptysis, blood in the stool, blood stasis, blood stasis and uterine bleeding and other blood-heat syndrome, water and fire burns, eczema and bloated.

Modern research shows that the mantle contains mainly a variety of tannins, triterpenoids and their glycosides, flavonoids, sugars and inorganic trace elements, including tannins 17~25%, saponins 2. 5~4% (mantle) Research Progress, Chinese Herbal Medicine, 1996, 27 (Supplement): 213; Chinese Medicine Chemistry, Shanghai People's Publishing House, 1975). The mantle promotes wound healing, bacteriostatic, anti-inflammatory, hemostasis, sedative, anti-inflammatory, anti-diarrhea, etc. (Modern Research and Application of Traditional Chinese Medicine) (Volume II), edited by Zheng Huzhan, Beijing, Xueyuan Press, 1997 , 10); Mantle extract has an inhibitory effect on UV-induced skin damage in rats (Forensic Medicine of Traditional Chinese Medicine, 2002, 24 (5): 299); Japanese Mimaki Y et al. reported that it was isolated from the root of mantle. Several ursane-type triterpenoid saponins have cytotoxic activity, ie, can inhibit HSC-2 (population bottom cancer) cells and HGF (hybridoma growth factor) [phytochemistry. 2001, 57 (5): 773-779]; The publication No. CN 1504478 A discloses the use of total saponin extract of mantle in raising white blood cells or/and platelets, indicating that mantle can be used for various reasons, especially leukopenia caused by chemoradiotherapy in cancer patients. And thrombocytopenia. Dang Chunlan et al studied the effects of mantle on the hemorheology of normal rabbits. The authors used a mathematical model to investigate the relationship between whole blood viscosity and hemostatic effect of mantle. It is believed that the hemostatic effect of mantle is related to the increase of whole blood viscosity. . (Dang Chunlan, Cheng Fangrong. The effect of mantle on blood rheology in rabbits. Chinese Journal of Medical Physics, 1997, 14 (3): 138~: 139) But so far, there is no original drug or ground for mantle The extracts or mantles of various active ingredients are used to treat and/or prevent anemia in raising red blood cells and hemoglobin.

1

Confirmation SUMMARY OF THE INVENTION

The technical solution of the present invention is to record: 7; the use of the ground and the extract, specifically, in the preparation of the elevated art magnetics

3⁄4 of Chinese medicine: HM^ is derived from the dried root of Sanguisorba officinalis L. or Longleaf 榆 Sanguisorba officinalis L. Var Longifolia (Bert) YU et Li.

The mantle is a mantle native powder or a water/organic solvent extract.

The mantle organic solvent extract is one of an organic solvent such as methanol, ethanol, ethyl acetate, n-butanol or diethyl ether, or a mixed solvent extract of any two or more kinds of solvents in any ratio.

Wherein, the aqueous or organic solvent mantle extract contains total saponins of mantle, and the weight percentage thereof is 10 98%. It is preferably 40 to 98%.

Wherein, the total saponin of the mantle contains a ursane-type triterpenoid saponin compound of the formula (I).

(I)

Wherein: is one of 11 and sputum; H, D-glucose, D-galactose, L-arabinose, L-rhamnose, D-xylose, D-glucuronic acid, L-galactosaldehyde One of acid, D-xylose glucose, acetyl-L-arabinose; one of OH, CH ₂ ; one of H, C 3⁄4; R _s is a species of H, D-glucose .

Wherein, the compound of the formula (I), 3⁄4 is H, R ₂ is α-L-arabinose, R ₃ is OH, is -0-glucose, ie 3-O- a -L-Arabic Glycosyl-19α-hydroxy-Uso-12--28-carboxylic acid-28-Ο-β-Ρ-glucosyl ester (ziyu- glycoside I, compound A).

The compound of the formula (I), wherein R _s is H, and R ₂ is α-Ο-L-arabinose, which is OH, that is, 19 α-hydroxyisob-2-ene-28-carboxylic acid- 3- a -0-L-arabinoside (ziyu- glycoside Π, compound Β)ο

The compound of the formula (I), Ri, R ₂ , is H, R ₃ is OH, and R ₅ is β-D-glucose, that is, 3 β, 19 α -dihydroxy-Uso-12--28 - Carboxylic acid -28-0-β-D-glucosyl (Compound C).

The compound of the formula (I), R ₂ , », R ₅ is H, and R ₃ is OH, that is, 3 β,19 α -dihydroxy-Uso-12--28-carboxylic acid (Compound D ).

The compound of the formula (I) is H, is a-L-arabinose, 3⁄4, 4 is simultaneously =CH ₂ , and R _s is β-D-glucose, that is, 3 β [( _α -1- Arabinose) Hydroxy] Uso- 12,19(29)-diene-28-carboxylic acid -28-0-β-indole-glucosyl (compound oxime).

a compound of the formula (I), 3⁄4, is Η, 3⁄4, 3⁄4 is OH, is - glucose, 艮P is 2α, 3α, 19α-trihydroxy-Uso-12--28-carboxylate Acid -28-0-β-D-glucosyl ester (Compound F).

Further, the present invention provides the use of the total saponin of the mantle and the main active ingredient thereof, the ursane-type triterpenoid saponin compound of the formula (I), for the preparation of a medicament for increasing red blood cells and hemoglobin.

The present invention provides a pharmaceutical composition which is water or organic of the original medicinal material of the mantle, or the medicinal material of the mantle. The solvent extract, or the total saponin of the mantle or the ursane-type triterpenoid saponin compound of the formula (I) is an active ingredient, and a preparation prepared by adding a pharmaceutically acceptable adjuvant or auxiliary component.

The preparation is an oral preparation or an injection.

The invention also provides a preparation method of the total saponin of the mantle, which comprises the following steps:

a. After properly pulverizing the original medicinal materials, extract with water, methanol or ethanol at room temperature or with heat, filter, filter off the dregs, and concentrate the filtrate under normal pressure or reduced pressure, and let stand for use;

b. The a step extraction concentrate is adsorbed by a polyamide column, and then eluted with 30 to 90% ethanol, and the eluate is recovered under reduced pressure, and then repeatedly adsorbed on the polyamide column for 1 to 3 times to obtain an extract. The total glycosides of the mantle are obtained by vacuum or freeze-drying.

The invention also provides a preparation method of the pharmaceutical composition, which comprises the following steps:

a. The ground medicinal material is appropriately pulverized, and then extracted with water, methanol or ethanol at room temperature or heat for 1 to 3 times, each time for 1 to 3 hours; filtered, the drug residue is filtered off, and the filtrate is concentrated under normal pressure or reduced pressure. To a certain concentration, stand still; b. The a step extraction concentrate is adsorbed by a polyamide column, and then eluted with 30~90% ethanol. After the eluate is recovered under reduced pressure, the polyamide column can be repeatedly adsorbed 1~ 3 times, an extract, a vacuum or freeze-dried to obtain a dry powder, and then added pharmaceutically-used excipients or auxiliary ingredients to prepare a pharmaceutically-acceptable oral preparation or injection;

Or an effective amount of a ursane-type triterpenoid saponin compound of the formula (I) is added, and a pharmaceutically acceptable adjuvant or auxiliary ingredient is used to prepare an oral preparation or an injection which is commonly used.

Among them, the extraction solvent is preferably a 50 to 90% ethanol solution. The amount of the ethanol solution is 6 to 12 times the weight of the medicinal material, and 8 times is preferred. The eluent is 60 to 90% ethanol, preferably 70%. The dry powder prepared above is a pale yellow solid substance with a slight bitter taste, and the total saponin contained therein is 10 to 98% of the total weight.

The present invention also provides the use of the pharmaceutical composition for the preparation of a medicament for increasing the action of red blood cells and hemoglobin.

The ursane-type triterpenoid saponin compound of the formula (I) and the above preferred compounds A, B, C:, D, E, F can be derived from the natural plant mantle (Sanguisorba officinalis L), Long-leaf mantle Longifrfia ( Bettol.)Y3⁄4.et Li), Sanguisorba alpina Beg., Ilex corunta Lindl. 3⁄4 young leaves ( Kuding tea), Mao Dongqing (Hex pubescens Hook.et.Arn. ), isolated from Echinops grijisu root, mazus miqueU, Hex crenata, etc.; or chemically synthesized. Specific chemical extraction and separation methods can be carried out according to literature methods (The structures of glycosides and aglycone of Sanguisorbae Radix, Chem. Pharm. BuU. 1971, 19(8): 1700-1707; Triterpene glycosides from the roots of Sanguisorba O cinals, Phytochemistry. 2001, 57(5), 773-779 _; Separation, identification and determination of saponins in mantle. Chinese herbal medicine, 2003, 34(5): 397-399)

The present invention finds that the Chinese medicine mantle and its extract, the total saponin of the mantle or the ursin-type triterpenoid saponin compound of the formula (I) have an effect of increasing red blood cells and hemoglobin, on ⁶ °Co-γ rays, chemical drugs. The mice caused by anemia and hemorrhagic anemia have significant therapeutic effects.

The present invention also provides a pharmaceutical preparation comprising the traditional Chinese medicine mantle and its extract, the total saponin of the mantle or the ursane-type triterpenoid saponin compound of the formula (I) as an effective medicinal ingredient, which can be used for raising red blood cells And hemoglobin, which treats and/or prevents various anemias, especially anemia caused by hypofunction or failure of bone marrow hematopoiesis.

The above traditional Chinese medicine mantle and its extract, the total saponin of the mantle or the ursane-type triterpenoid saponin compound of the formula (Ϊ) are mixed with the pharmaceutically acceptable auxiliary additive component according to the corresponding conventional pharmaceutical preparation method. A drug that raises red blood cells and hemoglobin can be prepared. For example, after mixing with commonly used auxiliary additives such as disintegrators, excipients, lubricants, binders, fillers, and the like which are acceptable in an oral preparation, they can be made into tablets according to a conventional operation method and process. a drug in the form of a solid oral preparation such as a pill, a capsule or a plurality of corresponding sustained release agents, controlled release agents, etc.; and a surfactant such as a conventional solubilizer, emulsifier, wetting agent, foaming or defoaming agent, etc. After mixing a release agent, a preservative, a stabilizer, a flavoring agent, a thickener, etc., according to a conventional method, an oral preparation of a liquid preparation such as a liquid preparation, a syrup or an oral liquid can be prepared; and a conventional suspension agent The stabilizer, the dispersing agent or the reagent for adjusting the osmotic pressure of the solution is mixed, and is configured as an injection solution according to a corresponding conventional method.

The invention also provides a pharmaceutical composition comprising a traditional Chinese medicine mantle and an extract thereof, a total saponin of the mantle or a ursane-type triterpenoid saponin compound of the formula 0), or a traditional Chinese medicine mantle and an extract thereof, the mantle The total saponin or the ursane-type triterpenoid saponin compound of the formula 0) is combined with other western or traditional Chinese medicines having the effects of increasing red blood cells and hemoglobin.

According to the present invention, the above-mentioned Chinese medicine mantle and its extract, or the total saponin of the mantle or the ursin-type triterpenoid saponin compound of the formula (I), or the Chinese medicinal mantle and its extract, or the mantle may be used. a total preparation of a saponin or a ursane-type triterpenoid saponin compound of the formula (I) in combination with other western or traditional Chinese medicines having elevated red blood cells and hemoglobin, and prepared into an oral preparation or an injection according to a conventional preparation technique. .

Advantageous Effects of the Invention: Since the Chinese medicine mantle and its extract, the total saponin of the mantle or the ursane-type triterpenoid saponin compound of the formula (I) have an obvious effect of increasing red blood cells and hemoglobin, it can be used for the preparation of the treatment. And/or prevention of various anemias, especially pharmaceutical preparations caused by anemia of bone marrow hematopoietic function or failure, such as P preparations or injections. It has the characteristics of remarkable curative effect, low price and convenient taking.

Obviously, the roots of this article can be modified, replaced or altered in various other forms without departing from the basic technical idea of the present invention.

The above content of the present invention will be further described in detail below by way of specific embodiments in the form of embodiments. However, the scope of the above-mentioned subject matter of the present invention should not be construed as being limited to the following examples, and the technology implemented based on the above-mentioned contents of the present invention is within the scope of the present invention.

detailed description

Example 1 Preparation of Mantle Original Drugs

Take the traditional Chinese medicine mantle, that is, the dried root of the Sanguisorba officinalis L. or the Sanguisorba officinalis L. Var Longifolia (Bert) Yu et Li., after crushing, sieving, and making the particle size. For 60~120 destination 榆 fine powder, fine powder is filled into capsules, sterilized, ready to serve.

Example 2 Preparation of Mantle Total Saponin Extract 1

1000g of mantle, after pulverization, add 8L of 70% ethanol solution, heat and reflux to extract 1. 5 hours, filter, filter residue and add 6L of 70% ethanol solution, heat and reflux for 1 hour, and filter. The filtrate was combined twice, concentrated to a certain concentration, and allowed to stand overnight. The polyamide resin was packed in a column, and the above extract was adsorbed through a polyamide column, and then the column was eluted with 70% ethanol, and the eluate was collected. The eluate was concentrated under reduced pressure, and ethanol was recovered to obtain an extract. The obtained extract was dissolved by heating with water, and then passed through a polyamide column twice to obtain an extract, which was vacuum dried to obtain 24 g of a dry powder, which was a mantle extract 1. Take the mantle extract lg, accurately weighed, placed in a 250ml flask, add 20ml each of 7% hydrochloric acid solution and absolute ethanol, heated to reflux for 2h in water bath, the temperature of the water bath is controlled at 90~95 ° C, filtered, the filtrate is on the water bath Was evaporated, the residue was transferred to a separatory funnel with 15 ml of distilled water, and extracted with ethyl acetate for 5 times, 15 ml each time. The ethyl acetate extract was evaporated to dryness in water, and the residue was dissolved in methanol. Methods (Zhao Yuxin, Li Manling. Determination of oleanolic acid and ursolic acid in traditional Chinese medicines and preparations by high performance liquid chromatography-evaporative light scattering detection. Journal of Pharmaceutical Analysis, 2004, 24 (2): 167~170) The total saponin content of the mantle in the total saponin extract 1 of the mantle is 74% of the total weight.

Example 3 Preparation of Mantle Total Saponin Extract 2

Burnet 1000 _g, was added 80% ethanol solution 8L crushed, heated at reflux for 1.5 hours extraction, filtration, The residue was further added to an 80% ethanol solution and heated under reflux for 1 hour, and filtered. The filtrate was combined twice, concentrated to a certain concentration, and allowed to stand overnight. ^ Resin packed column, the above extract was adsorbed through a polyamide column, and then 3⁄4 JC solution was selected with 75% ethanol, and the eluate was concentrated under reduced pressure to recover ethanol to obtain an extract. The obtained extract was heated with water to heat the acylate 3⁄4 times, and the extract was vacuum dried to obtain 19 g of dry powder, which was the mantle extract 2, which was speculated by the method described in Example 2, wherein the total saponin content of the mantle accounted for 55% of the total weight. .

Example 4 ^ 3 Preparation

The ground mantle 1000g, 13⁄4 amount of 70% ethanol was heated and refluxed 3 times for 2 hours each time. The extracts were combined 3 times, allowed to cool, filtered, and the ethanol was recovered to obtain an extract. The extract was dissolved in 3 times the amount of water, extracted 3 times with petroleum ether, 300 ml each time, and the fat-soluble impurities were removed. The aqueous layer is extracted with 600 ml of n-butanol in three portions, the aqueous layer is discarded, and n-butanol is recovered to dryness under reduced pressure to obtain 53 g of dried saponin. The crude saponin 15 is dissolved in 3 amounts of water, and saturated with salt. 5克的粉末状状。 . For the mantle extract 3, it was detected by the method described in Example 2, wherein the total saponin content of the mantle was 96% of the total weight.

In the case that the mechanism of the extraction process of the upper sputum is unchanged, the same extraction effect can be achieved by correspondingly changing and replacing the organic solvent for extraction, and an extract of different saponins content of different mantle is obtained.

Example 5 Extraction, Separation and Structure Confirmation of Ursane-type Triterpenoid Saponins Compounds A, B and C

Extraction and Separation: The mantle medicinal material was purchased from Chengdu Chinese Medicine Company and was identified as Sanguisorba officinalis L. Take 20kg of medicinal herbs, slice, and extract twice with 8 times 70% ethanol for 1 hour, filter, recover ethanol, add water to 0. 3g crude drug / ml, refrigerate for 12 hours, take the supernatant through the large hole Adsorption resin. The mixture was washed with water until the Molish reaction was negative, and then eluted with 70% ethanol. The eluate was collected, ethanol was recovered, and concentrated to obtain a concentrated extract. The concentrated extract was thoroughly stirred by adding 25 L of hot water, and the precipitate was filtered off by suction (about 4 k _g , and TLC was mainly detected as enamel). The filtrate was extracted three times with an equal volume of ethyl acetate and n-butanol, respectively, and then concentrated to dryness under reduced pressure to give 300 g of ethyl acetate extract, and n-butanol extract 320 s. 19 fractions were eluted with petroleum ether: acetone = 20:1 to 2:1 gradient, and the 16th fraction was allowed to stand in methanol to obtain a solid; the solid was separated by a silica gel column, petroleum ether: acetone =5:1 elution, white crushed crystals were precipitated at 34 to 36 minutes to obtain compound B 20 mg _; the obtained n-butanol extract was separated by a silica gel column, and the initial eluent was chloroform:methanol = 10:1 Then, elute with a gradient of 6:1, 4:1, 2:1. When the gradient is 4:1, a solid precipitates, and after repeated dissolution with methanol, it is precipitated, and finally a white powder of 852 mg, that is, compound A is obtained; the obtained n-butanol extract is separated by an R _P- 18 column, and methanol and water are started. (40% methanol) eluent, then eluted with 50%, 70%, 90%, 100% methanol, mixed solids in 70% methanol, and then separated with Sephadex LH-20 to give 38 mg of compound :.

Structure confirmation: The carbon and hydrogen spectra of the obtained compounds A, B, and C are consistent with those reported in the literature (Dong-Liang Cheng > Xiao-Ping Cao, Pomolic acid derivatives from the root of Sanguisorba officinalis, Phytochemistry, 1992). , 31 (4) : 1317—1320), ie: Compound A is 30 −〖0-( 1 -arabinose) 1-19 1 -hydroxyloxa-12-ene-28-carboxylic acid-28--glucose ester (ziyu- glycoside I ); Compound B is 3 β -[0-( α -L-arabinose)]-W α -hydroxyiso-12-ene- 28-carboxylic acid (ziyu- glycoside II )-, compound C 3⁄4 3 β , 19 α -Dihydroxy Uso-12-ene-28-carboxylic acid -28-β-indole-glucose. The ¹ H NMR and ¹³ C NMR data of the compounds A, B, and C are shown in Table 1 below.

The above-mentioned extraction and separation of the ursane triterpenoid saponin compounds A, Bv, C and the determination of the structure are the basis of the quality control of the drug of the present invention, and the quality of the drug can be determined.

Example 6 Preparation of tablets

Composition of tablets: Mantle extract 0.5g

HPMC drawing 30g

Lactose 69g

Magnesium stearate 0.5g

Total 100g

Mantle extract (Examples 2, 3 or 4, different extracts can be made into tablets of different specifications within the scope of the extract),

Lactose is mixed, and the wet granules are made of 75% ethanol as a binder. After 22 mesh sieve, 50 Ό dry for 3 hours, 22 mesh sieve granules, add magnesium stearate and mix and compress to make 1000 tablets, each piece 0.1 g. For patients with anemia, oral, 3 times a day, 2 tablets each time.

Example 7 Preparation of Capsules

Capsule composition: Mantle extract 0.5g

Starch 99g

Magnesium stearate 0.5g

Total 100g

Mantle extract (Examples 2, 3 or 4, different extracts can be prepared in different sizes within the scope of the extract), starch, magnesium stearate, and capsules, 1000 capsules in total , 0.1g per capsule. For patients with anemia, oral, 3 times a day, 2 capsules each time.

Example 8 Preparation of Injection

Injection composition: Mantle total saponin 0.5g

Tween 80 10ml

Sodium chloride 8g

Mantle total saponin extract (Examples 2, 3 or 4, different extracts can be prepared in different sizes within the scope of the extract), adding 10% sodium carbonate to adjust the pH to 7.0~7.5, refrigerated filtration , add Tween-80, add water for injection to lOOml, filter, dispense, potting, steam sterilization at 100 °C, 30 min.

Example 9 Preparation of Injection

Injection composition: Compound A 0.2g

Tween 80 10ml

Sodium chloride 8g

Add Compound 10 to 10% sodium carbonate to adjust the pH to 7.0~7.5, refrigerate and filter, add Tween-80, add water for injection to 1 Qing 0 leg!, filter, dispense, potting, WMTC steam sterilization, 30min That is.

Example 10 Preparation of Compound Tablets

Preparation of a compound preparation using the mantle extract, taking 0.5 _{g of the} mantle extract (Examples 2, 3 or 4, in the Extract the range of different extracts tablets may be prepared by different specifications), the total astragalosides 200g, according to conventional methods, add ingredients to give tablets _called? Per tablet / granules 200 _m g, for anemia, oral, 3 times a day, 1 tablet each time or 1 month.

The beneficial effects of the present invention are demonstrated by pharmacodynamic tests below.

Test Example 1: Effects of mantle native powder, total mantle saponin, compounds A, B, and C on anemia caused by cyclophosphamide in mice

Kunming mice, male and female, weighing 22~24g, were randomly divided into 9 groups according to body weight: normal control group, model group, mantle primitive drug group, mantle saponin high, medium and low dose group, compound A, Group B, Group C. Except the normal control group, each group was intraperitoneally injected with cyclophosphamide 100 mg/kg for 3 consecutive days. After injection of cyclophosphoryl group, the sputum was sputum for 10 consecutive days, ^Μ ^ ^Α^Μ 10 eyes to obtain the number of red blood cells in Jfiff mice. (RBC), blood: protein content (HGB) and hematocrit (HCT). Caesium; E Table 2: Mantle native powder, mantle saponins, compounds A, B, C on anthocyanin-induced anemia mice RBC, HGB and

The influence of HCT (X soil s)

Group dose sample number RBC HGB HCT

(mg/kg) (only) (10 ^U L) (g L) (%)

Normal control group - 12 10. 00±0. 74 153.51 ±9. 72 40.54+1. 84 Model control group - 10 9.05±0.86 ^AA 136.84±7. ^{β7 ΑΑ} 36.84±2. 29 ^AA ^mantle native Qing 2000 12 9.78±0.62* 149.25±10.74* 37.55±3.61 Total amount of high-alloy group 10 12 11.27±3.21** 161.33±12.59** 42.86±5.61** Manganin total dose group 5 12 10.05±1.08* 150.46±14.76 39.97±4.37* Mantle saponin low dose group 1 12 9.77±1.23* 145.02±11.37* 38.42±3.31 Food group A. 0. 24 12 11.55±0.99** 164.89±18.29** 43.91±4.06" Compound B group 0. 24 12 9.93±0.?8* 147.4 ±12.97* 36.07±3.12 Compound C group 0. 24 12 10.11±1.02* 145.06±12.56* 37.12±3.77 Note: Compared with normal control group: ^A P<0.05, P<0.01

Comparison with model control group - * P < 0.05, '* P < 0.01

It can be seen from the results in Table 2 that the number of red blood cells and the hemoglobin content decreased after 3 days of continuous injection of cyclophosphamide in mice, which was significantly different from that in the normal group, indicating that the model was successful. Compared with the model group, the number of red blood cells, hemoglobin, and hematocrit of the mantle were significantly increased, and the difference was statistically significant. It is indicated that the mantle native powder, the total saponin of the mantle, and the compounds A, B and C have the effect of treating anemia caused by chemotherapy drugs.

Test Example 2: Effects of mantle primary powder, mantle saponins, compounds A, B, and C on anemia induced by ^6e Co-Y ray in mice

Kunming mice, male and female, weighing 22~24 _g , were randomly divided into 9 groups according to body weight: normal control group, model group, mantle primary drug group, mantle saponin high, medium and low dose group, compound A , B, C groups. Except normal Outside the control group, the other groups of mice received ^6e Go-Y radiation for one-time exposure, the source skin distance was 5m, and the total absorption of each mouse was 750cGy. The above irradiation was carried out in Chengdu Fenggui. The next day after irradiation, the stomach was administered. For 10 consecutive days, B venous plexus was taken for blood, and the RBC:, HGB and HCT values of each mouse were determined. The results are shown in Table 3.

Table 3: Mantle native powder, mantle saponin extract 1, compound A, B, C to ^6β )-γ-ray-induced anemia mice

Impact of BBC, HGB and HCT (X±s)

Group dose sample number RBC HGB HCT

(mg/kg) (only) (10 ¹² /L) (g L) (%) Normal pair of cars 1 12 10.23 ± 0.65 146.67 ± 10.40 39.39 ± 2.81 Model control group - 10 8.49 ± 0.72" 128.90 ± 9.89 ^AA 33.78 ±2.18 ^AA mantle thick drug group 2000 12 10.26±1.58* 150.25±25.12** 37,55*7.52* Ground dose group% 12 11.18±1.39** 159.17±31.24** 40.15i8.25* Mantle saponin medium Group 5 12 10.11±1.93* 148.29±29.37** 37.53⁄4t6.95* Mantle total saponin low dose group 1 12 9.21±1.45* 136.27±21.54* 36.55±7.11* Compound A group 0.24 12 12.36±1.55** 165.05± 33.42** 41.56±6.74** Compound B group 0.24 12 10.12±0.96* 150.35±28.17* 36.72±9.86 Compound C group 0.24 12 9.56 o±1.25* 147.31±26.19* 36.22±9.77

Note: Compared with the normal group: ^A Ρ<0.05, "P<0.01

Compared with the model group: * P < 0.05, P < 0.01

From the results in Table 3, it can be seen that the number of red blood cells and hemoglobin content of the mice after receiving ^10D Co-Y-rays for one-time exposure 10 is at a low level, which is significantly different from that of normal control mice (P<0.01). , indicating that ^6e Co-Y-ray induced mouse anemia model is established. Compared with the model group, the number of red blood cells, hemoglobin and hematocrit in the mantle were significantly increased compared with the model group, and the difference was statistically significant. , indicating that the mantle native powder, the total saponin of the mantle, and the compounds A, B, and C have the effect of treating radiation-induced anemia.

Test Example 3: Total saponins of mantle, compounds A, B, and C on RBC, HGB, and HCT in hemorrhagic anemia mice

Eighty mice, weighing 18~22 _g , were randomly divided into 8 groups, normal control group, model group, high saponin, medium and low dose group, and compound A, B and C groups. Except the normal control group, the other groups were treated with 0.5 ml of blood in the eyeball (strictly control the amount of blood transfusion), and administered intraperitoneally 24 hours later for 7 consecutive days. After 1 hour of the last administration, the eyeballs were taken for blood determination of RBC, HGB and HCT. The results are shown in Table 4.

Table 4 Effects of total saponins, compounds A, B, and C on RBC, HGB, and HCT in mice with hemorrhagic anemia (X±s) Group doses RBC HGB HCT

C mg/kg) (g) (%) Normal control group - 10 10.72 ± 1.02 169.25 ± 9.37 41.71 ± 2.42 Model control group - 10 9.00 ± 1.37" 157.38 ± 10.96 ^{Α Δ} 35.21 ± 6.34 ^{Δ Α}榆Total saponin extract 1 high 2 10 11.51±1.13** 172.93±12.68** 43.22±5.75** Mantle total saponin extract 1 1 10 10.78±8».96'"' 165.21±10.S5* 41.12± 3.16* ^A mantle saponin extract 1 low 0. 2 10 9.42±0.77* 160.73±9.61 37.73±2.06 Compound A group 0. 24 10 11.58±2.15"180.22±11.65" ^A 42.35±3.57 Compound B group 0. 24 10 10.06±1.96* 168.76±11.57 ^ft * 39.78±3.15* Compound C group 0. 24 10 9.78±0.86* 165.23±10.22* 38.97±3.58* Note: Compared with the normal group: ^A P<0.05 ^A P<0.01

_; 3⁄43⁄4»3⁄4 ·3⁄4 ^ίΚ05,'· ^; ' ** . P<0.01

From the results in Table 4, i can be seen. Compared with the normal group, the RBC, HGB and HCT of the mice were significantly reduced after the modeling, and the difference was significant (P<0.01), indicating that the modeling was successful; After total saponins and compounds A, B, and C, compared with the model group, the number of red blood cells, hemoglobin, and hematocrit in the mice increased significantly, and the difference was statistically significant, indicating that the total saponins and compounds A were , B, C have a significant therapeutic effect on hemorrhagic anemia. The above ^ ^ injection ^ test shows that the hair of the present invention is as good as ittc i red 3⁄4 white ^.

The irf is not the same as the 6-volume group and the different pharmacodynamic effects test. The drug extract of the present invention increases the red blood cells and white: it has obvious effect, indicating that the drug of the present invention adopts oral sputum and injection of southern medicine, and has obvious high The role of cells and hemoglobin, and its effect is enhanced with the dose of the drug. It can be treated with 3⁄4 and / ^f, which is made up of low blood pressure and low blood pressure. 3⁄4 «, and the quality is stable, controllable Strong.

Claims

Claim

1. The use of a traditional Chinese medicine mantle for preparing a drug for raising red blood cells and hemoglobin.

2. The use according to claim 1, characterized in that the traditional Chinese medicine mantle is derived from the Sanguisorba officinalis L. or the Sanguisorba officinalis L. Var Longifolia (Bert) YU et Li. Dry roots.

3. Use according to claim 1 or 2, characterized in that the mantle is a mantle native powder.

Use according to claim 1 or 2, characterized in that the mantle is water or an organic solvent extract.

The use according to claim 4, wherein the mantle organic solvent extract is one or any two or more of methanol, ethanol, ethyl acetate, n-butanol and diethyl ether organic solvents. A solvent mixture of solvents in any ratio.

The use according to claim 4, characterized in that the mantle water or organic solvent extract contains total mantle saponins in a weight percentage of 10 to 98%.

The use according to claim 6, characterized in that the mantle water or organic solvent extract contains mantle total saponins in a weight percentage of 40 to 98%.

The use according to claim 6 or 7, wherein the total saponin of the mantle comprises a ursane-type triterpenoid saponin compound of the formula (I):

Wherein: is one of 11, OH; H, D-glucose, D-galactose, L-arabinose, L-rhamnose, D-xylose, D-glucuronic acid, L-galactosaldehyde One of acid, D-xylose-D-glucose, acetyl-L-arabinose; one of OH, CH ₂ ; one of H, C 3⁄4; R _s is H, D-glucose One kind.

9. Use of total saponins of mantle in the preparation of a medicament for increasing red blood cells and hemoglobin.

10. Use of a ursane-type triterpenoid saponin compound of the formula (I) for the preparation of a medicament for increasing red blood cells and hemoglobin.

11. A composition for raising red blood cells and hemoglobin, which is an aqueous or organic solvent extract of a mantle or a medicinal material of the mantle, or a total saponin of the mantle or ursane of the formula (I) A triterpenoid saponin compound is an active ingredient, and a preparation prepared by adding a pharmaceutically acceptable adjuvant or auxiliary ingredient.

The pharmaceutical composition according to claim 11, wherein the preparation is an oral preparation or an injection.

13. A method of preparing a total saponin of mantle comprising the steps of:

a. After properly pulverizing the original medicinal material, extract it with water, methanol or ethanol at room temperature or with heat, and filter. Filtered dregs, filtered heteroaryl Qu difficult £ Hfe prayer _{_Zen,;;;}

B, will be _a very natural rejection Me ^^ recorded miieii% alcohol eluate recovered under reduced pressure, the adsorption column may be repeated polyamide 1~3 times, hunting extract, burnet total drying or by freeze-blank 3⁄43⁄4 ^{; :} -; ^? ? ^: ; ·τ '^ - - ^:^ :- ■·■ .

A method for preparing a pharmaceutical composition according to claim 11 or 12, which comprises the steps of: a. pulverizing the mantle medicinal material, using water, methanol or ethanol, extracting at room temperature or heating, filtering , filtering off the dregs. The filtrate is concentrated under normal pressure or reduced pressure, and allowed to stand still;

b. The a step extraction concentrate is adsorbed by a polyamine column, and then eluted with 30-90% ethanol. After the eluate is recovered under reduced pressure, the oxime amide is adsorbed 1 to 3 times to obtain an extract. The saponin total saponin is obtained by vacuum or freeze-drying, and then a pharmaceutically-used auxiliary or auxiliary ingredient is added to prepare a pharmaceutically acceptable oral preparation or injection;

Or an effective amount of the ursane-type triterpenoid saponin compound of the formula (I), or a pharmaceutically-acceptable adjuvant or an auxiliary preparation, or a pharmaceutically-acceptable preparation or injection.