CN100496522C - Medicinal composition containing propolis, preparation method and use thereof - Google Patents

Abstract

The invention relates to a drug component with propolis. Wherein, it comprises: 5-60 propolis, 5-60 anoelica sinensis, and 5-60 chuanxiong rhizome; its production comprises that: selecting 5-60 anoelica sinensis and 5-60 chuanxiong rhizome, extracting the volatile oil; extracting the drug slag, adding water, extracting the eccentric liquid, concentrating to obtain the concentrated liquid; drying the concentrated liquid; processing the propolis, volatile oil and the concentrated dried ones, to obtain the final product. The invention can improve the immunity.

Description

A kind of pharmaceutical composition of propolis-containing, preparation method and its usage

Technical field

The present invention relates to a kind of pharmaceutical composition, preparation method and its usage, relate to specifically a kind of by propolis, Radix Angelicae Sinensis, Rhizoma Chuanxiong compatibility, through extracting new pharmaceutical composition, the Its Preparation Method And Use that effective site prepares.

Background technology

Dry secretions for Apidae insecticide apis mellifera Linnaeus Apis mellifera L..Nature and flavor: hardship, suffering, cold.But anti-inflammation, adjusting immunity, antioxidation, acceleration organization healing.Bibliographical information, propolis extract have the effect that strengthens mouse cell immunologic function and humoral immune function; Have and strengthen macrophage phagocytosis of mice and the active effect of NK cells in mice, show propolis can strengthen immune function of mice (Shao Yan, Pu Jinbao. propolis is regulated immune function of mice experimentation [J]. Zhejiang combination of Chinese and Western medicine magazine, 2005,15 (2): 90; Li Shuhua, in Xiao Hong, Yu Yingjun, etc. propolis is to immunologic hypofunction model mouse immune function influence [J]. Chinese medicine journal, 2001,29 (3): 39)

Its nature and flavor hardship of propolis, cold need be joined hot using warming therapy thing to press down the property of its bitter cold; Radix Angelicae Sinensis, sweet suffering is warm in nature, and matter is moistened and is bored with; Rhizoma Chuanxiong, Xin Wen and dry.Radix Angelicae Sinensis relatively nourishes blood and blood, and Rhizoma Chuanxiong is partial to the promoting the circulation of blood blood that looses, and same usefulness is matched in the every mutual-assistance of two medicines, can strengthen blood circulation promoting and blood stasis dispelling, nourish blood and the merit of blood.In addition, two medicines are moisturized suitable, but the profit Rhizoma Chuanxiong (processed) suffering of Radix Angelicae Sinensis is dry, and the hot dry anti-again Radix Angelicae Sinensis of Rhizoma Chuanxiong is greasy, and blood stasis dispelling and wasting QI-blood not nourish blood and the stagnation of QI is stopped up in not hyperamization.Full side's compatibility can press down the destitute property of propolis, can make again return, the property of rhizome of chuanxiong coordinates mutually, the impairment of QI-blood not.

The characteristics of this compositions: propolis be equipped with Radix Angelicae Sinensis, Rhizoma Chuanxiong etc. have invigorate blood circulation, the Chinese medicine of circulation of qi promoting effect, make this product have eliminating evil, blood stasis dispelling, and can adjust the effect of healthy energy in the body again, thus reach the effect of human body immunity improving power, be different from the existing product in market.Therefore, compare with the existing propolis series products in market, this product has unique advantage.

Summary of the invention

The object of the present invention is to provide a kind of new pharmaceutical composition, a kind of specifically by propolis, Radix Angelicae Sinensis, Rhizoma Chuanxiong compatibility, through extracting the new pharmaceutical composition that effective site prepares.

Another object of the present invention is to provide the preparation method of aforementioned pharmaceutical compositions.

A further object of the present invention is to provide the purposes of a kind of aforementioned pharmaceutical compositions aspect preparation raising body immunity medicine.

Purpose of the present invention can reach by following measure:

A kind of pharmaceutical composition of propolis-containing is characterized in that mainly being made up of the raw material of following weight portion: propolis 5～60, Radix Angelicae Sinensis 5～60, Rhizoma Chuanxiong 5～60; Preferably: propolis 25, Radix Angelicae Sinensis 50, Rhizoma Chuanxiong 25.

Above-mentioned pharmaceutical composition can be made any appropriate drug preparation by this area conventional method after adding adjuvant.As powder, oral liquid, granule, tablet or capsule, be preferably soft capsule, granule, oral liquid, effervescent tablet, dispersible tablet, chewable tablet, oral cavity disintegration tablet or liquid hard capsule, most preferably be soft capsule.

A kind of method for preparing above-mentioned pharmaceutical composition comprises the steps:

A, get Radix Angelicae Sinensis 5～60 weight portions, Rhizoma Chuanxiong 5～60 weight portions extract volatile oil, and volatile oil is standby;

B, get the medicinal residues that extract behind the volatile oil, extracting in water, extracting solution is centrifugal, concentrate, concentrated solution;

C, get step B and extract the concentrated solution obtain, drying, dry thing, standby;

D, get material and propolis that steps A, C obtain, through post processing, the finished product preparation of compositions;

The purposes of aforementioned pharmaceutical compositions aspect preparation raising body immunity medicine.

Purpose of the present invention specifically can reach in following measure:

Pharmaceutical composition involved in the present invention, its crude drug comprises propolis, Radix Angelicae Sinensis, Rhizoma Chuanxiong, its weight proportion is: propolis 5～60, Radix Angelicae Sinensis 5～60, Rhizoma Chuanxiong 5～60.When preparing said composition, elder generation extracts the volatile oil in Radix Angelicae Sinensis, the Rhizoma Chuanxiong, and residue through extracting, concentrating, gets extract again, with volatile oil and extract difference or mixed processing, obtains the various dosage forms of this pharmaceutical composition.This pharmaceutical composition has the effect of anti-human body immunity improving power.

Propolis is the dry secretions of Apidae insecticide apis mellifera Linnaeus Apis mellifera L.; Radix Angelicae Sinensis is the root of umbelliferae angelica Angelica sinensis (Oliv.) Diels; Rhizoma Chuanxiong is the root of samphire Rhizoma Chuanxiong Li gusticum chuanxion Hort..

The said medicine combination is extracted the volatile oil of Radix Angelicae Sinensis, Rhizoma Chuanxiong earlier in the preparation, and the residue after volatile oil is extracted is again through extracting, extracting solution concentrates, get spissated extracting solution,, be prepared into the various dosage forms of this pharmaceutical composition volatile oil and spissated extracting solution difference or mixed processing.Its detailed process comprises the steps:

A, get Radix Angelicae Sinensis 5～60 weight portions, Rhizoma Chuanxiong 5～60 weight portions are with CO ₂Supercritical extraction extraction volatile oil wherein, extracting pressure 10～50MPa resolves pressure 4～6MPa, 20 ℃～45 ℃ of extraction temperature, extraction time 2h～8h gets volatile oil, and residue is standby.

B, get the Radix Angelicae Sinensis, the Rhizoma Chuanxiong medicinal residues that extract behind the volatile oil, extracting in water 1～3 time, each 0.5～3 hour, high speed centrifugation when extracting solution is concentrated into every ml and contains 1～4g medical material, it is 1.05～1.35 (60 ℃) that centrifugal liquid continues to be concentrated into relative density, gets the concentrated solution of extracting solution.

C, get the concentrated solution of step B gained, drying, the dry extract of medicine.

D, with steps A carry volatile oil and the extract of step C, propolis mix, through post processing, the finished product preparation of compositions.

The dosage form of finished product preparation is an acceptable forms on the pharmaceutics among E, the above-mentioned steps D, as soft capsule, tablet, capsule, granule, oral liquid, effervescent tablet, dispersible tablet, powder, chewable tablet, oral cavity disintegration tablet, liquid hard capsule etc., be preferably capsule, soft capsule; Most preferably be soft capsule.

The specification of the most preferred medicinal composition soft capsule of the present invention is: 0.5g/ grain, 0.6g/ grain, 1g/ grain.Oral, one time 2,3 times on the one, or follow the doctor's advice.

Pharmaceutical composition of the present invention has the effect that improves immunity, can be used for treating immunologic hypofunction disease.

In this compositions propolis be equipped with Radix Angelicae Sinensis, Rhizoma Chuanxiong etc. have invigorate blood circulation, the Chinese medicine of circulation of qi promoting effect, make this product have eliminating evil, blood stasis dispelling, and can adjust the effect of healthy energy in the body again, thereby reach the effect of human body immunity improving power, be different from the existing product in market.Therefore, compare with the existing propolis series products in market, this product has unique advantage.

The specific embodiment

The present invention is further illustrated below in conjunction with embodiment.

Embodiment 1

Get propolis 2500g, pulverize, standby; Get Radix Angelicae Sinensis 5000g again, anti-Rhizoma Chuanxiong 2500g puts CO ₂The extraction of supercritical fluid extraction device, temperature: 30 ℃, pressure: 30MPa extracted 3 hours, and the 6MPa decompress(ion) gets volatile oil, and is standby.Medicinal residues behind the extraction volatile oil add 10 times of water gagings, decoct 2 times, decoct 1 hour at every turn, filter, and merging filtrate is concentrated into per 1 milliliter of medicinal liquid that contains the 3g crude drug in whole, high speed centrifugation; Centrifugal liquid continues to be concentrated into relative density 1.15～1.20 (60 ℃), and spray drying gets the dry fine powder of pale brown color, mixes with propolis powder, above-mentioned volatile oil, add an amount of dextrin, starch, magnesium stearate, to total amount 4000g, mixing is granulated, tabletting, encapsulation, promptly.

Embodiment 2

Get propolis 2500g, pulverize, standby; Get Radix Angelicae Sinensis 5000g again, anti-Rhizoma Chuanxiong 2500g puts CO ₂The extraction of supercritical fluid extraction device, temperature: 30 ℃, pressure: 30MPa extracted 3 hours, and the 6MPa decompress(ion) gets volatile oil, and is standby.Medicinal residues behind the extraction volatile oil add 10 times of water gagings, decoct 2 times, decoct 1 hour at every turn, filter, and merging filtrate is concentrated into per 1 milliliter of medicinal liquid that contains the 3g crude drug in whole, high speed centrifugation; Centrifugal liquid continues to be concentrated into relative density 1.15～1.20 (60 ℃), and spray drying gets the dry fine powder of pale brown color, mixes with propolis powder, above-mentioned volatile oil, adds an amount of dextrin, starch, and to total amount 4000g, mixing fills in Capsules, promptly.

Embodiment 3

Get propolis 2500g, pulverize, standby; Get Radix Angelicae Sinensis 5000g again, anti-Rhizoma Chuanxiong 2500g puts CO ₂The extraction of supercritical fluid extraction device, temperature: 30 ℃, pressure: 30MPa extracted 3 hours, and the 6MPa decompress(ion) gets volatile oil, and is standby.Medicinal residues behind the extraction volatile oil add 10 times of water gagings, decoct 2 times, decoct 1 hour at every turn, filter, and merging filtrate is concentrated into per 1 milliliter of medicinal liquid that contains the 3g crude drug in whole, high speed centrifugation; Centrifugal liquid continues to be concentrated into relative density 1.15～1.20 (60 ℃), spray drying, get the dry fine powder of pale brown color, mix with propolis powder, above-mentioned volatile oil, add an amount of Polyethylene Glycol-400, to total amount 6000g, in 50～60 ℃ of mixings, make the soft capsule of 0.6g/ grain with the soft capsule preparation technology, promptly.

The pharmacodynamic experiment research of this drug regimen:

With the soft capsule of embodiment 3 gained, per os gives the mice various dose, and (1800mg/kg.bw) 1 month, other established the 0mg/kg.bw group and replaces given the test agent with edible oil for 300mg/kg.bw, 600mg/kg.bw.

Experimental technique is undertaken by the enhancing immunity functional check method of " health food check and assessment technique standard " (version in 2003):

1), ConA inducing mouse splenocyte transformation experiment---mtt assay

Each dosage treated animal continuous irrigation stomach is after 1 month, the cervical vertebra dislocation method is put to death animal, and the aseptic spleen of getting places the little plate that fills an amount of aseptic Hank ' s liquid, grind spleen, make the individual cells suspension, filter, use Hank ' s liquid to wash 2 times through 200 eye mesh screens, each centrifugal 10min (1000r/min), then with cell suspension in 1mL RPMI640 complete culture solution, the blue dyeing counting viable count (all more than 95%) of platform phenol is 3 * 10 with RPMI640 complete culture solution adjustment cell concentration ⁶Individual/mL.Divide two holes to add in 24 well culture plates cell suspending liquid, every hole 1mL, a hole adds 75 μ L ConA liquid, and 37 ℃, 5%CO are put in contrast in another hole ₂Cultivate 72h in the incubator.Cultivate and finish preceding 4h, supernatant 0.7mL is inhaled in every hole gently, adds the RPMI RPMI-1640 that 0.7mL does not contain calf serum, adds MTT (5mg/mL) 50 μ L/ holes simultaneously, continues to cultivate 4h.After cultivating end, every hole adds 1mL acid isopropyl alcohol, and the piping and druming mixing dissolves purple crystal fully.Lysate is moved in 96 well culture plates, under wavelength 570nm, measure each sample cell optical density value with microplate reader.

By table 1 result as seen, 1800mg/kg.bw group adds the ConA hole and is significantly higher than the 0mg/kg.bw group with the difference that does not add ConA hole absorbance.

The influence that table 1 soft capsule transforms the inductive mouse spleen lymphocyte of ConA

Dosage (mg/kg.bw)	Number of animals (only)	Add ConA hole and the difference that does not add ConA hole absorbance	The P value ( ±s)
				0	10	0.010±0.007
300	10	0.021±0.013	0.145
				600	10	0.017±0.014	0.448
1800	10	0.028±0.016	0.006

2), serum hemolysin is measured---half hemolysis value (HC ₅₀)

Each dosage treated animal continuous irrigation stomach prepared the SRBC suspension of 2% (v/v) after 1 month, and every Mus lumbar injection 0.2mL carries out immunity, extractd eyeball behind the 4d and got blood in centrifuge tube, placed 1h, and the centrifugal 10min of 2000r/min separates and also collects serum.After 200 times of dilutions of serum, the optical density value during by method of inspection working sample pipe and SRBC HD50.The amount of hemolysin is with half hemolysis value (HC ₅₀) expression.

By table 2 result as seen, 1800mg/kg.bw group HC ₅₀Be significantly higher than the 0mg/kg.bw group.

Table 2 soft capsule is to mice HC ₅₀Influence (

± s)

Dosage (mg/kg.bw)	Number of animals (only)	HC 50	The P value ( ±s)
				0	10	94±31
300	10	96±25	0.995
				600	10	104±28	0.709
1800	10	126±23	0.031

3), Turnover of Mouse Peritoneal Macrophages is engulfed chicken red blood cell experiment---half intracorporal method

Each dosage treated animal continuous irrigation stomach 1 month, the chicken red blood cell suspension of preparation 20% (v/v), every Mus lumbar injection 1mL, interval 30min, mice is put to death in the cervical vertebra dislocation, it is faced upward the position be fixed on the Mus plate, and abdominal skin is cut off in the center, inject normal saline 2mL through the abdominal cavity, rotate Mus plate 1min, sucking-off abdominal cavity washing liquid 1mL then, average mark drips on 2 microscope slides, put into the enamel box that is lined with wet gauze, 37 ℃ of constant incubators of dislocation are hatched 30min, then, and rinsing in normal saline, dry, fix with 1:1 acetone methanol solution, 4% (v/v) Giemsa-phosphate buffer dyeing 3min, the rinsing of reuse distilled water is dried, mounting, light microscopic is observed down.

By table 3 result as seen, 600mg/kg.bw, 1800mg/kg.bw group Turnover of Mouse Peritoneal Macrophages is engulfed chicken red blood cell percentage rate and phagocytic index and all is higher than the 0mg/kg.bw group.

Dosage (mg/kg.bw)	Number of animals (only)	Phagocytic percentage (%)	The P value ( ±s)	Phagocytic index	The P value ( ±s)
						0	10	15.9±2.8		0.19±0.03
300	10	19.4±2.7	0.149	0.23±0.04	0.307
						600	10	24.2±5.7	0.000	0.28±0.09	0.007
1800	10	23.0±5.5	0.002	0.28±0.09	0.006

4), the NK cytoactive is measured---the determination of lactate dehydrogenase method

Each dosage treated animal continuous irrigation stomach is after 1 month, the cervical vertebra dislocation method is put to death Mus down, the aseptic spleen of getting, place the little plate that fills an amount of aseptic Hank ' s liquid, grind spleen, make the individual cells suspension, filter through 200 eye mesh screens, use Hank ' s liquid to wash 2 times, each centrifugal 10min (1000r/min) abandons supernatant cytoplasm is upspring, and adds the 0.5mL aquesterilisa 20 seconds, add 0.5mL2 times of Hank ' s liquid and 8mL Hank ' s liquid after the splitting erythrocyte again, centrifugal 10min (1000r/min), it is resuspended to contain 10% calf serum RPMI, 1640 complete culture solutions with 1mL, with 1% glacial acetic acid dilution back counting, the blue dyeing counting viable count (all more than 95%) of platform phenol, adjusting cell concentration with RPMI 1640 complete culture solutions is 2 * 10 ⁷Individual/mL.

24h washes target cell (YAC-1 cell) cultivations of going down to posterity 3 times with Hank ' s liquid before using before the experiment, is 4 * 10 with RPMI1640 complete culture solution adjustment cell concentration ⁵Individual/mL.Getting each 100 μ l of YAC-1 cell and splenocyte (imitating target than 50:1) adds in U type 96 well culture plates, YAC-1 cell nature release aperture adds YAC-1 cell and each 100 μ l of culture fluid, the maximum release aperture of YAC-1 cell adds YAC-1 cell and each 100 μ L of 1%NP40, above-mentioned every three parallel holes of all establishing are in 37 ℃, 5%CO ₂Cultivate 4h in the incubator, then with 96 well culture plates with the centrifugal 5min of 1500r/min, draw at the bottom of the supernatant 100 μ L horizontalizations in 96 well culture plates in every hole, add LDH substrate liquid 100 μ L simultaneously, reaction 8min, every hole adds the HCl 30 μ L of 1mol/L, measures optical density value at microplate reader 490nm place.

By table 4 result as seen, 600mg/kg.bw group NK cells in mice activity is significantly higher than the 0mg/kg.bw group.

Table 4 soft capsule to the active influence of NK cells in mice (

± s)

Dosage (mg/kg.bw)	Number of animals (only)	NK cytoactive (%)	The P value ( ±s)
				0	10	58.6±16.2
300	10	85.3±25.8	0.068
				600	10	94.8±40.5	0.014
1800	10	80.1±30.5	0.181

By above experiment as seen, this compositions has the enhancing immunity function.

This drug regimen toxicity test:

(1) soft capsule of this compositions is oral through female, male mice, and acute toxicity dosage belongs to non-toxic type greater than 22.4g/kg.bw;

(2) soft capsule of this compositions gave rat 30 days continuously, and dosage is respectively 1800mg/kg.bw, 3900mg/kg.bw, and 6000mg/kg.bw, the animal subject ordinary circumstance is good, all no abnormal change of body weight, food utilization, organ weights, organ coefficient; Hematological indices and biochemical indicator result show that every index is all in normal range; Each internal organs histopathologic examination there is no relevant pathological change.

Claims (9)

1, a kind of pharmaceutical composition of propolis-containing is characterized in that being made up of the raw material of following weight portion:

Propolis 5～60, Radix Angelicae Sinensis 5～60, Rhizoma Chuanxiong 5～60.

2, pharmaceutical composition according to claim 1 is characterized in that the weight proportion of propolis, Radix Angelicae Sinensis and Rhizoma Chuanxiong is:

Propolis 25, Radix Angelicae Sinensis 50, Rhizoma Chuanxiong 25.

3, a kind of pharmaceutical composition can be made any appropriate drug preparation by this area conventional method after it is characterized in that the described pharmaceutical composition of claim 1 added adjuvant.

4, pharmaceutical composition according to claim 3 is characterized in that described preparation is powder, oral liquid, granule, tablet or capsule.

5, pharmaceutical composition according to claim 4 is characterized in that described preparation is soft capsule, granule, oral liquid, effervescent tablet, dispersible tablet, chewable tablet, oral cavity disintegration tablet or liquid hard capsule.

6, pharmaceutical composition according to claim 5 is characterized in that described preparation is a soft capsule.

7, a kind of method for preparing the described pharmaceutical composition of claim 3 comprises the steps:

A, get Radix Angelicae Sinensis 5～60 weight portions, Rhizoma Chuanxiong 5～60 weight portions extract volatile oil, and volatile oil is standby;

B, get the medicinal residues that extract behind the volatile oil, extracting in water, extracting solution is centrifugal, concentrate, concentrated solution;

C, get step B and extract the concentrated solution obtain, drying, dry thing, standby;

D, get material and propolis that steps A, C obtain, through post processing, the finished product preparation of compositions.

8, preparation method according to claim 7 is characterized in that comprising the steps:

A, get Radix Angelicae Sinensis 5～60 weight portions, Rhizoma Chuanxiong 5～60 weight portions, with CO2 supercritical extraction extraction volatile oil wherein, extracting pressure 10～50MPa resolves pressure 4～6MPa, 20 ℃～45 ℃ of extraction temperature, extraction time 2h～8h, volatile oil, residue is standby;

B, get the Radix Angelicae Sinensis, the Rhizoma Chuanxiong medicinal residues that extract behind the volatile oil, extracting in water 1～3 time, each 0.5～3 hour, high speed centrifugation when extracting solution is concentrated into every ml and contains 1～4g medical material, when centrifugal liquid continues to be concentrated into relative density and is 60 ℃ 1.05～1.35, the concentrated solution of extracting solution;

C, get the concentrated solution of step B gained, drying, the dry extract of medicine;

D, with steps A carry volatile oil and the extract of step C, propolis mix, through post processing, the finished product preparation of compositions.

9, the purposes of the described pharmaceutical composition of claim 1 aspect preparation raising body immunity medicine.