CN1739528A - Application of monomeric alkaloid extracted from Hubei fritillary bulb in preparing medicine for relieving cough, eliminating phlegm and relieving asthma - Google Patents

Abstract

The present invention discloses the application of monomeric alkaloid extracted from Hubei fritillary bulb in preparing medicine for relieving cough, eliminating phlegm and relieving asthma. The comparison of Hubei fritillary bulb alkaloid, monomeric alkaloid glycoside and total alkaloid in relieving cough, eliminating phlegm and relieving asthma shows that under the condition of the same dosage, Hubei peimisine has higher effect, and under the condition of the same toxicity, peininine has higher effect.

Description

The application of the monosomic alkali that extracts in the Hupeh Fritillary Bulb in preparation relieving cough and expelling phlegm suppressing panting calming medicine

Technical field:

The present invention relates to Chinese medicine extract and application thereof, the application of several monosomic alkali that are specifically related to from Hupeh Fritillary Bulb, extract in preparation relieving cough and expelling phlegm suppressing panting calming medicine.

Background technology:

Hupeh Fritillary Bulb is the dry bulb of Liliaceae Fritillaria plant Hupeh Fritillary Bulb (Fritillaria Hupehensis Hsiao et K.C.Hsia), and lung, heart channel are gone in little hardship, cold, have the effect of clearing heat and moistening lung, preventing phlegm from forming and stopping coughing and eliminating stagnation.Hupeh Fritillary Bulb was recorded by Chinese crude drug standard promulgated by the ministries or commissions of the Central Government in 1991, had now recorded in the Pharmacopoeia of the People's Republic of China (version in 2000), developed into the second largest main flow kind that is only second to Bulbus Fritillariae Thunbergii.Aspect chemical constitution study, from the bulb of Hupeh Fritillary Bulb (be called for short lake shellfish) isolation identification 11 kinds of alkaloids: lake shellfish ester (Hupeheninetate), peimine (peimine), verticinone (peiminine), hupehenine (hupehenine), hupehenirine (hupehenirine), hupehenizine (hupehenizine), hupeheninoside (hupeheninoside), hupehemonoside (hupehemonoside), hupehenidine (hupehenidine), Hupehenisine (hupehenisine) and Ebeisine (ebeiensine).In addition, we produce from Hubei to separate in north, Hubei Province Bulbus Fritillariae Uninbracteatae and north, pale reddish brown Hubei Province Bulbus Fritillariae Uninbracteatae the Fritillaria plant and obtain ebeinine (ebeinine), ebeinone alkaloids such as (ebeinone).

Modern pharmacology studies show that [referring to Xiong Wei, Guo Xiaoling, He Jialang. the preliminary study of Hupeh Fritillary Bulb pharmacological action (J). Chinese herbal medicine, 1986,17 (3): 115-118.] the Hupeh Fritillary Bulb ethanol extraction has obvious relexation to several kind of smooth muscle cells, Cavia porcellus there is antiasthmatic effect, can improves mice tolerance normobaric hypoxia ability; Total alkaloids is its biological activity effective site; Total alkaloids has antitussive, eliminates the phlegm, antiasthmatic effect, and its alkaloid and biological activity be than Bulbus Fritillariae Cirrhosae, Bulbus Fritillariae Thunbergii height, but toxicity is also big than Bulbus Fritillariae Cirrhosae, Bulbus Fritillariae Thunbergii.

Cough, expectorant, to breathe heavily be the common sympton of respiratory system disease, especially bronchial asthma.At present slow in the medicine progress in this field, still there is not desirable medicine.Common cough suppressing panting calming medicine such as Herba Ephedrae preparation, untoward reaction is many, as hypertension, has a sleepless night, has a headache, reaches tachylaxis.Western medicine such as aminophylline are relievingd asthma only and are not had antitussive, phlegm-dispelling functions, and can produce the tolerance phenomenon.Therefore, the cough suppressing panting calming medicine of a kind of safe, stable, high curative effect of excavation, no toleration has significance from the traditional medical treasure-house of motherland.

Summary of the invention:

The monomer of the objective of the invention is in several monosomic alkali in the Bulbus Fritillariae Uninbracteatae are produced in Hubei, to filter out antitussive, eliminate the phlegm, antiasthmatic activity is good, exploitation become a kind of natural, efficient, low toxicity antitussive, eliminate the phlegm, suppressing panting calming medicine.

The objective of the invention is to reach by following measure:

Produce from Hubei that extraction separation obtains alkaloid and glycoalkaloid monomer the Bulbus Fritillariae Uninbracteatae, respectively under the Isodose condition and under the equal toxicity condition to alkaloid and glycoalkaloid monomer carried out antitussive, the experiment of eliminating the phlegm, relieving asthma.

To producing the Bulbus Fritillariae Uninbracteatae alkaloid and the glycoalkaloid monomer has carried out antitussive, the experiment of eliminating the phlegm, relieving asthma in Hubei, and with the total alkaloids comparison, draw as drawing a conclusion: (1) is under the Isodose condition: ebeinine, hupeheninoside show significant antitussive effect; Ebeinine, Ebeisine show significant expectorant activity; Ebeinone, verticinone, Ebeisine tool be antiasthmatic effect preferably; (2) under equal toxicity condition, hupehenirine, verticinone, hupeheninoside, hupehenine are being equivalent to each alkaloid sample LD ₅₀1/10 amount the time show significant antitussive effect; Hupeheninoside, verticinone dosage are LD ₅₀1/10, peimine, hupehenine, Ebeisine dosage are LD ₅₀Showed significant expectorant activity at 1/5 o'clock; Peimine, verticinone, Ebeisine are being equivalent to each alkaloid sample LD ₅₀3/20 when amount show the antiasthmatic activity of highly significant.

The specific embodiment:

Embodiment 1: each monomer antitussive under the Isodose condition, eliminate the phlegm, the antiasthmatic activity experiment

The preparation of sample: accurately take by weighing each monomer 3mg with ten thousand/balance, be settled to 10ml with 0.1%CMC-Na solution, to the ultrasonic cleaner ultrasonic 10 minutes, shake up, standby.

(1) antitussive experiment

Get 100 of body weight 20 ± 2g white mice, male and female half and half are divided into 10 groups at random, behind table 1i.g administration 1h, place in the inverted 500ml beaker, draw ammonia 0.15ml with the 1ml syringe and inject a cotton balls of putting in the beaker, observe the cough latent period and the number of times of coughing in 3 minutes.

Each the monomer gastric infusion mice antitussive effect experiment of table 1 Hupeh Fritillary Bulb

Animal dosage and approach cough number of times

Group cough latent period (second)

Number of elements (mg/kg) (in 3 minutes)

Negative 10 0 i.g 58.9 ± 14.1 13.1 ± 4.2

Positive 10 30 i.g 91.1 ± 25.2 ^*7.4 ± 2.4 ^*

Ebeinine 10 3 i.g 87.0 ± 30.7 ^*3.9 ± 1.9 ^*

Ebeinone 10 3 i.g 71.1 ± 18.6 15.2 ± 6.1

Peimine 10 3 i.g 76.2 ± 25.2 12.0 ± 5.7

Verticinone 10 3 i.g 65.5 ± 21.9 11.0 ± 4.2

Hupeheninoside 10 3 i.g 83.6 ± 24.4 ^*10.8 ± 5.1

Hupehenine 10 3 i.g 50.3 ± 13.9 15.3 ± 7.8

Ebeisine 10 3 i.g 62.1 ± 28.1 15.1 ± 5.8

Total alkali 10 3 i.g 98.7 ± 15.6 6.8 ± 1.7

Negative group: 0.1%CMC-Na solution; Positive group: codeine

Compare with feminine gender ^*P＜0.05 ^*P＜0.01

Ebeinine, hupehenine, total alkali group show significant antitussive effect, with feminine gender significant difference (p＜0.01-0.05) are arranged relatively.

(2) experiment of eliminating the phlegm

Get 80 of body weight 20 ± 2g mices, male and female half and half are divided into 10 groups at random, and water 12h is can't help in fasting before the experiment, by table 2i.g administration, 0.5h lumbar injection 0.5% phenol red solution 0.5ml after the administration puts to death mice (as far as possible not damaging trachea) after 30 minutes, face upward the position and be fixed on the operation plate, cut off neck center skin, separate trachea, No. 7 syringe needles that under larynx head polished insert 0.3cm in the trachea, draw 5%NaHCO with the fixing back of silk thread ligation with the 1ml syringe ₃Solution 0.5ml by syringe needle lavation respiratory tract 3 times (not stopping) back and forth at every turn, injects a color comparison tube with irrigating solution, draws 5%NaHCO again ₃Solution 0.5ml is lavation 3 times more as above, the 3rd same lavation again, shared washing liquid 1.5ml like this takes out and washes 9 times, merges eluate, with spectrophotometer in wavelength 546nm place survey A value.

Each the monomer gastric infusion mice phlegm-dispelling functions experiment of table 2 Hupeh Fritillary Bulb

The phenol red excretion amount of dosage and approach (ug/ml)

The group number of animals

(mg/kg) (X±SD)

Negative 10 0 i.g 0.0760 ± 0.03

Positive 10 25 i.g 0.1020 ± 0.03 ^*

Ebeinine 10 3 i.g 0.0994 ± 0.02 ^*

Ebeinone 10 3 i.g 0.0582 ± 0.03

Peimine 10 3 i.g 0.0610 ± 0.01

Verticinone 10 3 i.g 0.0650 ± 0.02

Hopeheninoside 10 3 i.g 0.0780 ± 0.03

Hupehenine 10 3 i.g 0.0800 ± 0.03

Ebeisine 10 3 i.g 0.0910 ± 0.03 ^*

Total alkali 10 3 i.g 0.0730 ± 0.03

Negative group: 0.1%CMC-Na solution; Positive group: guaifenesin

Compare with feminine gender ^*P＜0.05 ^*P＜0.01

Ebeinine, Ebeisine show significant expectorant activity, with the feminine gender ratio significant difference are arranged, ebeinine P＜0.01 wherein, and Ebeisine P＜0.05 all is better than total alkali.

(3) experiment of relievining asthma

Get the Cavia porcellus (Cavia porcellus childhood) of 170 ± 20g, male and female all can, measure one by one to draw and breathe heavily incubation period, select eligible.Cavia porcellus is put into 402 type ultrasound atomizer, spray into the equivalent mixed liquor 15 seconds of 2% acecoline and 0.4% histamine with the constant voltage of 53Kpa (400mmHg), keep a close eye on the reaction of Cavia porcellus, observe promptly to draw incubation period that the tic of panting property appears in Cavia porcellus in 6 minutes and breathe heavily incubation period (stopping to the time that the tic of panting property occurs) and the number of animals that tic takes place from spraying.As see Cavia porcellus and fall and to take out immediately, in order to avoid it is dead, and write down to draw and breathe heavily incubation period, then thought insensitive greater than 150 seconds incubation period and will not select for use if draw to breathe heavily, press body weight random packet with qualified Cavia porcellus (60) next day, every group 6,9 groups of Cavia porcelluss are treated test agent and matched group respectively, the i.g administration is drawn with similarity condition after 30 minutes and is breathed heavily and measure to draw and breathe heavily incubation period and the variation that the tic number of animals takes place.As draw and breathe heavily prolongation of latency, the number of animals of take place twitching reduces, and handles significance by statistics, can think that this medical instrument has antiasthmatic effect.

Each the monomer gastric infusion mice antiasthmatic effect experiment of table 3 Hupeh Fritillary Bulb

After the preceding medication of medication

Twitch in animal dosage and way

Group is drawn to breathe heavily to draw incubation period and is breathed heavily difference incubation period (second)

Number of elements footpath (mg/kg) number of animals

(second) (second)

Negative 60 34.2 ± 9.9 0/6 49.2 ± 16.8 15.0 ± 12.07

Positive 64 35.0 ± 9.56 6/6 76.7 ± 15.1 41.8 ± 17.0 ^*

Ebeinine 6 3.3 33.2 ± 9.1 6/6 47.5 ± 9.7 14.3 ± 7.9

Ebeinone 6 3.3 31.0 ± 9.4 6/6 78.7 ± 57.6 47.7 ± 49.9

Peimine 6 3.3 33.0 ± 6.7 6/6 68.0 ± 21.0 35.0 ± 26.0

Verticinone 6 3.3 32.8 ± 8.7 6/6 81.7 ± 57.8 48.8 ± 56.2

Hupeheninoside 6 3.3 33.7 ± 12.7 6/6 48.5 ± 21.5 14.8 ± 16.4

Hupehenine 6 3.3 33.7 ± 5.4 6/6 50.2 ± 15.7 16.5 ± 15.2

Ebeisine 6 3.3 33.0 ± 13.1 6/6 82.0 ± 42.4 49.0 ± 38.3

Total alkali 6 3.3 30.5 ± 3.6 6/6 52.7 ± 16.2 22.2 ± 13.8

Negative group: 0.1%CMC-Na solution; Positive group: ephedrine

Compare with feminine gender ^*P＜0.01

Draw that to breathe heavily the incubation period difference more approaching before the ebeinone, verticinone, Ebeisine administration with after the administration, difference is bigger, though with feminine gender than P＞0.05, difference obviously is longer than total alkaloids, may be the less reason of number of animals.

Conclusion:

Under the Isodose condition, the Antitussive and Expectorant Effect of each monomer gastric infusion of Bulbus Fritillariae Uninbracteatae to mice (3mg/kg) produced in Hubei, to the antiasthmatic effect of Cavia porcellus (3.3mg/kg):

(1) ebeinine, hupeheninoside show significant antitussive effect, with feminine gender significant difference (p＜0.01-0.05), and total alkali shows very strong toxicity under the Isodose condition are arranged relatively.

(2) ebeinine, Ebeisine show significant expectorant activity, with feminine gender significant difference are arranged relatively, ebeinine P＜0.01 wherein, and Ebeisine P＜0.05 all is better than total alkali.

(3) ebeinone, verticinone, Ebeisine tool antiasthmatic effect preferably draws and breathes heavily the incubation period difference and be longer than total alkaloids before the administration with after the administration.

Embodiment 2: each monomer antitussive under the equal toxicity condition, eliminate the phlegm, the antiasthmatic activity experiment

The preparation of sample: by pharmacology requirement preparation sample, mice (is equivalent to LD by every 0.2ml ₅₀1/5 the amount) the tail intravenously administrable; Cavia porcellus (is equivalent to LD by every 0.4ml ₅₀1/10 the amount) intraperitoneal injection; And transfer PH about 5 with dilute sulfuric acid, and shake up, standby.

(1) antitussive experiment

Get 100 of body weight 20 ± 2g white mice, male and female half and half are divided into 10 groups at random, behind table 4i.v (tail vein injection) administration 1h, place in the inverted 500ml beaker, draw ammonia 0.15ml with the 1ml syringe and inject a cotton balls of putting in the beaker, observe the cough latent period and the number of times of coughing in 3 minutes.

Each the monomer tail vein injection mice antitussive effect experiment of table 4 Hupeh Fritillary Bulb

Animal LD ₅₀Cough latent period cough number of times

Group dosage (mg/kg)

Number of elements (mg/kg) (second) (in 3 minutes)

Negative 10 0 58.3 ± 34.84 81.8 ± 31.16

Positive 10 4 75.6 ± 48.22 53.3 ± 28.30 ^*

Total alkali 10 1.3 13.17 77.8 ± 35.59 102.3 ± 59.33

Hupehenirine 10 5.2 51.89 100.6 ± 50.75 ^*74.6 ± 46.32

Peimine 10 1.5 14.29 93.7 ± 47.77 63.6 ± 40.07

Verticinone 10 1.3 13.08 46.4 ± 36.80 38.2 ± 22.46 ^*

Hupeheninoside 10 2.0 20.80 51.6 ± 25.29 53.3 ± 27.47 ^*

Hupehenine 10 4.2 41.84 57.0 ± 28.51 52.3 ± 29.50 ^*

Ebeisine 10 3.9 39.31 66.1 ± 33.60 55.4 ± 28.35

Negative group: water for injection; Positive group: codeine phosphate, i.g

Compare with feminine gender ^*: p＜0.05 ^*P＜0.01

Hupehenirine, verticinone, hupeheninoside, hupehenine are being equivalent to each alkaloid sample LD ₅₀1/10 amount the time show significant antitussive effect, be better than total alkali, with the feminine gender ratio significant difference (p＜0.01-0.05) is arranged.

(2) experiment of eliminating the phlegm

Get 80 of body weight 20 ± 2g mices, male and female half and half are divided into 10 groups at random, and water 12h is can't help in fasting before the experiment, by table 5i.g administration, 0.5h lumbar injection 0.5% phenol red solution 0.5ml after the administration puts to death mice (as far as possible not damaging trachea) after 30 minutes, face upward the position and be fixed on the operation plate, cut off neck center skin, separate trachea, No. 7 syringe needles that under larynx head polished insert 0.3cm in the trachea, draw 5%NaHCO with the fixing back of silk thread ligation with the 1ml syringe ₃Solution 0.5ml by syringe needle lavation respiratory tract 3 times (not stopping) back and forth at every turn, injects a color comparison tube with irrigating solution, draws 5%NaHCO again ₃Solution 0.5ml is lavation 3 times more as above, the 3rd same lavation again, shared washing liquid 1.5ml like this takes out and washes 9 times, merges eluate, with spectrophotometer in wavelength 546nm place survey OD value.

Each the monomer tail vein injection mice phlegm-dispelling functions experiment of table 5 Hupeh Fritillary Bulb

Animal dosage LD ₅₀Phenol red excretion amount (ug/ml)

Group

Number of elements (mg/kg) is X ± SD (mg/kg)

Negative 10 0 0.207 ± 0.039

Positive 10 500 0.273 ± 0.040 ^*

Total alkali 10 2.6 13.17 0.263 ± 0.040 ^*

Hupehenirine 10 10.4 51.89 0.215 ± 0.030

Peimine 10 3.0 14.29 0.242 ± 0.033 ^*

Verticinone 10 1.3 13.08 0.244 ± 0.032 ^*

Hupeheninoside 10 2.0 20.80 0.298 ± 0.032 ^{* *}

Hupehenine 10 8.4 41.84 0.271 ± 0.046 ^{* *}

Ebeisine 10 7.8 39.31 0.257 ± 0.031 ^*

Negative group: water for injection; Positive group: guaifenesin, subcutaneous injection

Compare with feminine gender ^*: p＜0.05 ^*P＜0.01 ^{* *}P＜0.001

(wherein hupeheninoside, verticinone dosage are LD for peimine, verticinone, hupeheninoside, hupehenine, Ebeisine ₅₀1/10, peimine, hupehenine, Ebeisine dosage are LD ₅₀1/5) show with the feminine gender ratio significant difference is arranged by significant expectorant activity, wherein hupeheninoside, hupehenine p＜0.001 are better than total alkali, peimine, verticinone, Ebeisine p＜0.05.

(3) experiment of relievining asthma

Get the Cavia porcellus (Cavia porcellus childhood) of 170 ± 20g, male and female all can, measure one by one to draw and breathe heavily incubation period, select eligible.Cavia porcellus is put into 402 type ultrasound atomizer, spray into the equivalent mixed liquor 15 seconds of 2% acecoline and 0.4% histamine with the constant voltage of 53Kpa (400mmHg), keep a close eye on the reaction of Cavia porcellus, observe promptly to draw incubation period that the tic of panting property appears in Cavia porcellus in 6 minutes and breathe heavily incubation period (stopping to the time that the tic of panting property occurs) and the number of animals that tic takes place from spraying.As see Cavia porcellus and fall and to take out immediately, in order to avoid it is dead, and write down to draw and breathe heavily incubation period, then thought insensitive greater than 150 seconds incubation period and will not select for use if draw to breathe heavily, press body weight random packet with qualified Cavia porcellus (72) next day, every group 8,9 groups of Cavia porcelluss are treated test agent and matched group respectively, the i.g administration is drawn with similarity condition after 30 minutes and is breathed heavily and measure to draw and breathe heavily incubation period and the variation that the tic number of animals takes place.

Each the monomer guinea pig intraperitoneal injection antiasthmatic effect experiment of table 6 Hupeh Fritillary Bulb

After the preceding medication of medication

The animal dose difference

Group LD ₅₀Draw to breathe heavily to draw incubation period and breathe heavily incubation period

Number of elements (mg/kg) (second)

(mgg/kg) (second) (second)

Negative 80 58.09 ± 13.78 69.6 ± 31.83 16.91 ± 37.89

258.13±

Positive 8 125 77.75 ± 30.45 335.88 ± 68.24

70.25 ^**

Total alkali 8 1.98 13.17 53.14 ± 12.12 62.71 ± 28.63 9.57 ± 29.31

Hupehenirine 8 7.8 51.89 67.88 ± 20.30 66.00 ± 18.32 5.38 ± 23.83

Peimine 8 2.28 14.29 53.38 ± 11.73 151.88 ± 132.17 95.51 ± 121.79 ^*

Verticinone 8 2.28 13.08 62.63 ± 28.48 161.13 ± 128.98 98.51 ± 131.34 ^*

Hupeheninoside 8 1.98 20.80 53.25 ± 14.08 71.38 ± 37.40 18.12 ± 27.12

Hupehenine 8 3.06 41.84 58.88 ± 17.61 55.38 ± 25.63-4.13 ± 30.39

Ebeisine 8 5.88 39.31 49.25 ± 7.81 84.5 ± 39.51 41.63 ± 51.16 ^*

Negative group: water for injection; Positive group: aminophylline

Compare with feminine gender ^*P＜0.05 ^*P＜0.01

Peimine, verticinone, Ebeisine are being equivalent to each alkaloid sample LD ₅₀3/20 when amount show the antiasthmatic activity of highly significant, draw before and after the medication and breathe heavily the incubation period difference bigger increase is arranged, difference has significant difference (p＜0.05) with the feminine gender ratio.

Conclusion:

Under equal toxicity condition, each monomer is produced in the Bulbus Fritillariae Uninbracteatae to the Antitussive and Expectorant Effect of mouse tail vein administration in Hubei, to the antiasthmatic effect of guinea pig intraperitoneal injection:

(1) hupehenirine, verticinone, hopeheninoside, hupehenine are being equivalent to each alkaloid sample LD ₅₀1/10 amount the time show significant antitussive effect, be better than total alkali, with the feminine gender ratio significant difference (p＜0.01-0.05) is arranged.

(2) (wherein hupeheninoside, verticinone dosage are LD for peimine, verticinone, hopeheninoside, hupehenine, Ebeisine ₅₀1/10, peimine, hupehenine, Ebeisine dosage are LD ₅₀1/5) show with the feminine gender ratio significant difference is arranged by significant expectorant activity, wherein hupeheninoside, hupehenine p＜0.001 are better than total alkali, peimine, verticinone, Ebeisine p＜0.05.

(3) peimine, verticinone, Ebeisine show the antiasthmatic activity of highly significant when being equivalent to 3/20 amount of each alkaloid sample LD50, draw before and after the medication to breathe heavily the incubation period difference bigger increase is arranged, and difference has significant difference (p＜0.05) with the feminine gender ratio.

Claims (7)

1, the application in preparation relieving cough and expelling phlegm suppressing panting calming medicine of alkaloid that from Hupeh Fritillary Bulb, extracts or glycoalkaloid monomer.

2, described alkaloid that extracts from Hupeh Fritillary Bulb of claim 1 or glycoalkaloid monomer comprise peimine, verticinone, hupehenine, hupehenirine, hupehenizine, hupeheninoside, hupehemonoside, hupehenidine, Hupehenisine, ebeinine, ebeinone and Ebeisine.

3, the application of hupeheninoside in preparation relieving cough and expelling phlegm suppressing panting calming medicine.

4, ebeinine, Ebeisine, hupeheninoside, hupehenine, peimine or the verticinone application in the preparation expelling phlegm drugs.

5, Ebeisine prepares the application in the expelling phlegm drugs.

6, ebeinone, peimine, verticinone or the Ebeisine application in the preparation suppressing panting calming medicine.

7, verticinone or the Ebeisine application in the preparation suppressing panting calming medicine.