CN1125096A - Anatural mixture composed of higher primary aliphatic alcohols obtained from bee wax for the treatment of gastric and duodenal ulcers that also present antiinflamatory activity - Google Patents

Abstract

The present invention relates to a preparation method of new natural mixture composed of high-grade fatty primary alcohols of C22-38, specially C24-34, further specially C24, C26, C28, C30, C32 and C34. Those fatty primary alcohols are obtained from beeswax. ADVANTAGE-. The repeatability of the relative composition of ech kind of alcohol is good. Said natural mixture can be effectively used as active component for different medicament preparations to cure gastroduodenal ulcer and/or as protective agent for gastric and duodenal mucosa, and can display anti-inflammatory activity.

Description

By the natural mixture of forming from mellisic senior fat primary alcohol

The present invention relates to prepare a kind of new natural composition, it is by 22-38 carbon atoms, 24-34 carbon atoms particularly, and more especially 24,26,28,30, the senior fat primary alcohol of 32 and 34 carbon atoms is formed.The relative composition of every kind of alcohol of this mixture has high performance reproducibility between every batch.

The present invention relates generally to pharmaceutical industries, and particularly exploitation has the medicament preparation of specific performance, because they can be used for treating gastric duodenal ulcer, and can also be as antiinflammatory.

These preparations contain as active component, from 24-34 carbon atoms of peak wax, and particularly 24,26,28,30, the natural mixture of the linear senior fat primary alcohol of 32 and 34 carbon atoms (being called M.H.A.A.B.W. hereinafter).

As active component, the medicine with special pharmacological properties is rarely known by the people with the saturated linear senior fat primary alcohol of 24-34 carbon atoms for these.Only reported the natural mixture (EPA0 488 928) that derives from sugarcane wax in the past.

Since ancient times, used Apis to produce thing in the empiricism.Along with the development of analytical method, in many cases, differentiate that the active component in these products becomes possibility, this just makes people understand their some biological and pharmacokinetics effects.These products with therapeutic effect can exemplify peak royal jelly, Mel, pollen and Cera Flava.Trophism, scavenging action and other the pharmacy characteristic of considering them with and be used as the probability of skin mask, Cera Flava generally is used for medicine and cosmetics industry.Cera Flava mainly is made up of the chemical compound of following type: the saturated and undersaturated long chain hydrocarbon (55-75%) of-21 to 37 carbon atoms;-fatty acid ester (about 30%), it contains linear long-chain alcohol (16-30 carbon atoms); The free fatty of-16-30 carbon atoms (about 1-5%), and the acid in 18: 1 that account for acidic moiety 30%;-free long-chain alcohol (about 1-7%), what content was the abundantest is 30 carbon alcohol, octacosanol and 26 carbon alcohol;-other polar compound (about 1-3%).

Step of the present invention is based on the Cera Flava alkali metal and the saponified method of alkaline earth metal hydroxide concentrated solution homogeneous phase of fusing in advance, particularly use low-molecular-weight alkali metal alkaline earth metal hydroxide, more especially use sodium hydroxide, potassium hydroxide or calcium hydroxide.

The concentration of hydroxide solution must make the weight of corresponding hydroxide and pending mellisic weight ratio greater than 5%, is in particular 8-25%, is more particularly 15-25%.The saponification process kept 30 minutes at least, was in particular 2-5 hours.

To the solid that obtain place a solid-liquid extractor in this step, and optionally extract M.H.A.A.B.W. with the organic solvent of capacity, solvent is selected from C _3-8Ketone, C _6-9Hydrocarbon, C _1-5Alcohol, halogenated hydrocarbons and aromatic such as benzene and its derivant comprise their mixture.It is as follows to be used for solvents more of the present invention: acetone, methyl ethyl ketone, pentanone, hexanone, heptanone, 2-methylpentanone, ethanol, methanol, 2-propanol, butanols, the tert-butyl alcohol, pentane, hexane, heptane, octane, chloroform, 1,2-dichloroethanes, dichloromethane, trichloroethane, 1,2,3-trichloropropane, benzene, toluene, phenol, xylol and other.

Extraction was carried out 5-10 hours, and product is with above-mentioned solvent or their crystalline mixture then.Yield is about 30%, and the purity of product (M.H.A.A.B.W) is 80-98%, particularly 90-98%.

The product that so obtains in the present invention (M.H.A.A.B.W) is by C _24-34The mixture formed of senior fat primary alcohol.Its fusing point is 80.0-82.5 ℃.Here the method for from Cera Flava, obtaining M.H.A.A.B.W of Ti Chuing with before other method of report compared some advantages.Advantage is that preparation time is short, and another is the result who describes before the actual recovery [nearly 30% (weight)] of M.H.A.A.B.W is higher than, another advantage be products therefrom purity apparently higher than before report.In table 1, report the general composition of the quality and quantity of M.H.A.A.B.W, reported the composition of the quality and quantity of M.H.A.A.B.W in the table 2.

Table 1: the general composition of the quality and quantity of gained M.H.A.A.B.W

The percentage ratio of component in mixture

1-tetracosa carbon alcohol 9.0-15.0%

1-two ten six carbon alcohol 12.0-18.0%

1-octacosanol 13.0-20.0%

1-three ten carbon alcohol 20.0-30.0%

1-three ten two carbon alcohol 13.0-21.0%

1-three ten four carbon alcohols 1.5-3.5%

Table 2: the concrete composition of the quality and quantity of gained M.H.A.A.B.W

The percentage ratio of component in mixture

1-tetracosa carbon alcohol 12.5+/-1.0%

1-two ten six carbon alcohol 14.5+/-1.2%

1-octacosanol 16.5+/-2.0%

1-three ten carbon alcohol 24.6+/-1.6%

1-three ten two carbon alcohol 16.7+/-1.4%

1-three ten four carbon alcohols 2.3+/-0.5%

The daily dose that M.H.A.A.B.W is used for the treatment of various disease is 1-100mg/ days, and the most effective route of administration is oral, and dosage form is tablet and granule or capsule.Though consider the using method that the present invention recommends, this medicine can be through parenterai administration or topical.

Medicament preparation contains 0.5-25.0% active component M.H.A.A.B.W.By this M.H.A.A.B.W is obtained this dosage form with different excipient such as agglutinant, disintegrating agent, lubricant, Slider or filler mixture.These excipient comprise lactose, corn starch, sucrose, magnesium stearate, microcrystalline Cellulose, the crosslinked special-purpose Talcum of sodium carboxymethyl cellulose, gelatin, cellulose, acetyl phthalic acid ester, titanium dioxide, tablet, Polyethylene Glycol, polyvinylpyrrolidone and other.

One of purpose of the present invention be from Cera Flava begin to obtain, separation and purification be by (C _24-34The natural mixture formed of senior fat primary alcohol, particularly contain the natural mixture of the primary alconol of 24,26,28,30,32 and 34 carbon atoms.

Another object of the present invention is with the composition of low relatively dosage as the medicine Chinese medicine prescription of treatment gastric duodenal ulcer with these natural mixtures.Proved that this new M.H.A.A.B.W can obviously alleviate the gastric ulcer that is caused by aspirin, ethanol, the relevant medicine with other of indometacin (it can produce gastric ulcer in the treatment patient).In addition, this M.H.A.A.B.W also obviously alleviates duodenal ulcer.

Another object of the present invention is that exploitation contains the pharmaceutical formulation that M.H.A.A.B.W is divided into as activity, as anti-inflammatory agent, and Orally-administrable or parenteral route and topical.

At last, the global commerce of this M.H.A.A.B.W that obtains among the present invention is worth and recommends the active ingredient as pharmaceutical formulation, provable it be very safe and toleration good mixing thing, this is an important advantage.Acute, the inferior chronic and long term toxicity test result's that this point can obtain carrying out in rodent and rabbit support, the result shows the relevant toxicity of no medicine.In addition, it does not show any mutagenic action and teratogenesis in rodent.In the clinical trial for the treatment of, do not find any pair of effect with product of the present invention.

To illustrate in greater detail purpose of the present invention with reference to the following example, but not limit its scope.Embodiment 1

Get the Cera Flava that 1000g chops up,, add the 200g potassium hydroxide that is dissolved in the 150ml water 100-110 ℃ of fusings.Stir down and kept 30 minutes.The solid M.H.A.A.B.W that obtains extracts with heptane in solid-liquid extraction system.Extract is cooled to room temperature, uses the methyl ethyl ketone crystallization.Obtain 250g M.H.A.A.B.W, purity is 93.26%.The mixture fusing point is 81.0-82.5 ℃.The composition of the quality and quantity of the M.H.A.A.B.W that table 3 expression obtains with this step.

Table 3: the composition of the quality and quantity of gained M.H.A.A.B.W.

The percentage ratio of every kind of alcohol of component

1-tetracosa carbon alcohol 13.21

1-two ten six carbon alcohol 15.50

1-octacosanol 17.89

1-three ten carbon alcohol 26.03

1-three ten two carbon alcohol 18.05

1-three ten four carbon alcohols, 2.58 embodiment 2

Get the 2Kg Cera Flava,, add the 300g sodium hydroxide that is dissolved in the 200ml water 90-100 ℃ of fusings down.Stir down and kept 3 hours.The solid M.H.A.A.B.W that obtains tradition solid-liquid extraction system in alcohol extraction 12 hours.Extract is cooled to room temperature, with methylol recrystallization gained solid.Obtain 393g M.H.A.A.B.W, purity is 93.33%.The mixture fusing point is 80.5-82.0 ℃.The composition of the quality and quantity of the M.H.A.A.B.W that table 4 expression obtains with this step.

Table 4: the composition of the quality and quantity of gained M.H.A.A.B.W.

The percentage ratio of every kind of alcohol of component

1-tetracosa carbon alcohol 13.28

1-two ten six carbon alcohol 15.42

1-octacosanol 18.11

1-three ten carbon alcohol 26.10

1-three ten two carbon alcohol 17.73

1-three ten four carbon alcohols, 2.68 embodiment 3

Get the 50kg Cera Flava,, add the 12Kg calcium hydroxide that is dissolved in 101 water 100-120 ℃ of fusings.Stir down and kept 3.5 hours.The M.H.A.A.B.W that obtains uses chloroform extraction 12 hours in solid-liquid extraction equipment.Extract is cooled to room temperature, uses the heptane recrystallization subsequently.Obtain 14.5Kg M.H.A.A.B.W, purity is 93.77%.The mixture fusing point is 81.5-82.5 ℃.The composition of the quality and quantity of the M.H.A.A.B.W that table 5 expression obtains with this step.

The composition of the quality and quantity of table 5. gained M.H.A.A.B.W.

The percentage ratio of every kind of alcohol of component

1-tetracosa carbon alcohol 13.48

1-two ten six carbon alcohol 16.12

1-octacosanol 17.51

1-three ten carbon alcohol 26.55

1-three ten two carbon alcohol 17.73

1-three ten four carbon alcohols, 2.38 embodiment 4

Get the 50Kg Cera Flava,, add the 10.6Kg sodium hydroxide that is dissolved in the 7.5 L methanol 1/1 100-120 ℃ of fusings.Stir down and kept 4.5 hours.The M.H.A.A.B.W that obtains extracted 12 hours with 2-propanol in solid-liquid extraction equipment.Extract is cooled to room temperature, uses toluene: recrystallization in 1: 1 the mixture of chloroform.Obtain 9.3Kg M.H.A.A.B.W, purity is 93.36%.The mixture fusing point is 80.5-82.0 ℃.The composition of the quality and quantity of the M.H.A.A.B.W that table 6 expression obtains with this step.

Table 6: the composition of the quality and quantity of gained M.H.A.A.B.W.

The percentage ratio of every kind of alcohol of component

1-tetracosa carbon alcohol 13.33

1-two ten six carbon alcohol 15.54

1-octacosanol 17.02

1-three ten carbon alcohol 27.04

1-three ten two carbon alcohol 17.83

1-three ten four carbon alcohols, 2.60 embodiment 5

Two kinds of different medicament preparations have been prepared with this M.H.A.A.B.W as active component.The composition of every kind of preparation is shown in Table 7.Physics, chemistry and the physico-chemical property of considering M.H.A.A.B.W have prepared these preparations.

Table 7: with the medicament preparation of M.H.A.A.B.W as active component

Set of dispense goods 1 preparation 2

(％) (％)

M.H.A.A.B.W 5.0 15.0

Lactose 56.5 55.0

Corn starch 15.0 10.0

Polyvinylpyrrolidone 2.5 2.0

Cross-linking sodium carboxymethyl cellulose 5.0 4.0

Sucrose 5.0 4.0

Talcum 2.0 2.0

Magnesium stearate 1.5 1.0

Microcrystalline Cellulose 7.5 7.0 embodiment 6

Make the female Sprague Dawley rat of heavy 200-250g and the male guinea pig of heavy 300-400g under laboratory condition, adapt to 7 days.Rat is divided into different test group randomly.After the fasting 24 hours, inject the M.H.A.A.B.W that is suspended in 2%Tween 20/ water carrier in one group of rat peritoneum, and matched group is only injected the carrier (Tween20/ water) of equal volume.These processing are inducing ulcer to carry out in preceding 1 hour.What be used as the ulcer derivant is: NaOH (0.2mol/L), ethanol (60%), ASA (40mg/kg) and indometacin (50mg/kg) (indometacin is dissolved in 5% sodium bicarbonate), and by the administration of stomach feeding tube.Alcohol induced: carrier or 1,5,25,50 and the M.H.A.A.B.W administration of 100mg/kg after 1 hour, every rat is by oral 60% ethanol of stomach feeding tube (1ml/200g).After one hour, Mus is killed, gastric ulcer is carried out quantitatively.ASA induces: Cavia porcellus is divided into 4 test group, and one group in contrast, 3 groups of peritoneal injection M.H.A.A.B.W (5.25 and 100mg/kg).The carrier of matched group injection equal volume.After one hour the Mus gastric is given ASA (40mg/kg).After 2 hours, Mus is put to death, measure gastric ulcer.Indometacin is induced: rat is divided into 3 test group randomly: one for according to group, and two intraperitoneal give 25 and the M.H.A.A.B.W of 50mg/kg.Give indometacin by the stomach feeding tube after one hour, after 4 hours, put to death animal, gastric ulcer is quantitative.After each is induced in the example of ulcer, puts to death animal rapidly, take out stomach, open stomach from big crook, with the saline solution washing, measure the size of ulcer.The result damages by two different observers and measures with summation (mm) expression of damage.

In every example, treat the comparison of group and matched group with nonparametric U de Mann Whitney check.M.H.A.A.B.W is shown in respectively in the table 8,9 and 10 result of the effect of ethanol, ASA and the inductive gastric ulcer of indometacin.

Table 8:M.H.A.A.B.W is to the effect of the inductive gastric ulcer of ethanol (60%)

(X+/-DE)

Therapeutic dose (mg/kg) n ulcer size (mm)

Contrast 25 35,46+/-4,81

M.H.A.A.B.W. 1 10 35，28+/-9，13

5 10 4，11+/-1，57 ^＊＊＊

25 10 8，90+/-3，82 ^＊＊＊

100 10 9,35+/-3,88 ^{* *}P＜＜0,01; ^{* *}P＜＜0,001 (U de Mann Whitney check)

Table 9:M.H.A.A.B.W is to the effect of the inductive gastric ulcer of ASA (40mg/kg)

(X+/-ES)

Therapeutic dose (mg/kg) n ulcer size (mm)

Contrast 8 39,37+/-10,69

M.H.A.A.B.W. 5 6 38，00+/-13，90

25 8 9，50+/-1，72 ^＊

100 6 25,83+/-3,77 ^*P＜＜0,05 (U de Mann Whitney check)

Table 10:M.H.A.A.B.W is to the effect of the inductive gastric ulcer of indometacin (50mg/kg)

(X+/-ES)

Therapeutic dose (mg/kg) n ulcer size (mm)

Contrast 11 38,65+/-9.48

M.H.A.A.B.W. 25 11 17，50+/-4，38 ^＊

50 10 13，14+/-3，60 ^＊

^*P＜＜0,05 (U de Mann Whitney check).

M.H.A.A.B.W protects gastric mucosa (table 8,9 and 10) in the inductive ulcer model of ethanol, ASA and indometacin, obviously reduce the size of ulcer.Embodiment 7

The purpose of this experiment is to judge whether M.H.A.A.B.W depends on prostaglandin to the protective effect of anti-gastric-ulcer.Make the male Sprague Dawley Mus of heavy 150-200g adapt to laboratory condition 7 days with water that adds libitum and food, test fasting in preceding 48 hours.Animal is divided into 4 test group randomly.

Intraperitoneal is given the suspension of M.H.A.A.B.W in 2%Tween 20/ water, and the west is not dissolved in the hot water for fourth, and indometacin is dissolved in 5% sodium bicarbonate.Four test group following (n=10): 1) contrast (carrier); 2) and 3) give 25 and the M.H.A.A.B.W of 50mg/kg respectively, and 4) west of 25mg/kg is for fourth.After intraperitoneal is injected to animal, the subcutaneous indometacin 10mg/kg that gives.Give 60% ethanol after half an hour, put to death animal after one hour, take out stomach, open stomach, measure the ulcer size with 3 times of magnifieres from big crook.The result represents with millimeter.

In all examples, check relatively treatment group and matched group with nonparametric U de Mann Whitney.M.H.A.A.B.W the results are shown in the table 11 effect of ethanol and the inductive gastric ulcer of indometacin.

Table 11:M.H.A.A.B.W is to the effect of ethanol 100% and the inductive gastric ulcer of indometacin (10mg/kg)

Therapeutic dose (mg/kg) n ulcer (mm) suppression ratio (%)

Ethanol (60%) 10 35,3+2,95

Indometacin 10 0

Indometacin+ethanol 9 54,6+/-8,55

↑tween

M.H.A.A.B.W. 25 8 13，8+/-5，44 ^＊72，8

50 51 8，0+/-6，04 ^＊?64，5

Fourth 25 9 14,6+/-5,64 are not replaced in the west ^*71,2

^*P＜＜0,05 (U de Mann Whitney check)

From table, can see,, reduce, formerly take under the situation of indometacin by being like this by alcohol induced ulcer when the M.H.A.A.B.W of dosage shown in using or west during for fourth.Indometacin does not produce ulcer separately, but can increase the weight of alcoholic acid ulcer effect.

EXPERIMENTAL DESIGN is based on taking indometacin before the alcohol induced ulcer earlier, if chemical compound also keeps the antiulcer protective effect, then it does not rely on prostaglandin.The result shows that M.H.A.A.B.W still has the antiulcer effect after taking indometacin, can infer that this protective effect may not rely on prostaglandin.Embodiment 8

The foregoing description gained result hints that its effect is relevant or uncorrelated with the inductive ulcer of acid compound.In order to overcome the inhibitory action that gastric acid secretion is suppressed, in Pilorous ligation model, measure the effect of M.H.A.A.B.W to gastric acid secretion.

Make the female Sprague Dawley Mus of heavy 150-200g adapt to laboratory condition 15 days with water that adds libitum and food.Animal is divided into the different tests group randomly, in ligation fasting in preceding 48 hours.Intraperitoneal is given the suspension of M.H.A.A.B.W in 2%Tween20/ water.

The west is not dissolved in the hot water for fourth, by the administration identical with M.H.A.A.B.W.Animal is divided into 6 test group: 1) contrast (carrier); 2), 3) and 4) give 25,50 and the M.H.A.A.B.W of 100mg/kg, 5 respectively) and 6) (positive control) give respectively 25 and the west of 50mg/kg for fourth.

Rat with etherization and from abdominal incision, is taken out stomach, with silk thread ligation Pilorous.Intraperitoneal is given this chemical compound immediately then, and subcutaneous injection 3ml normal saline.After four hours, put to death animal, take out complete stomach, vise at the place at esophagus.The gastric thing is at 3000min ^-1Descended centrifugal 10 minutes, then measurement volumes.With 0.1 mol/L NaOH is the acidity that indicator is measured gastric juice with phenolphthalein.Check relatively treatment group and matched group result with nonparametric U de Mann Whitney.M.H.A.A.B.W is shown in Table 12 the effect of stomach acidity in the Pilorons ligation.

Table 12: M.H.A.A.B.W is to the influence of stomach acidity in Pilorous ligation model

Therapeutic dose η gastric juice volume acidity

(mg/kg) (mL) (meq?H ⁺/mL)

Contrast 12 8,18+/-0,51 0,109+/-0,002 M.H.A.A.B.W. 25 10 5,00+/-0,58 ^*0,09+/-0,01

50 11 5，51+/-0，54 ^＊ 0，104+/-0，006

100 10 5,4 9+/-0,65 ^*0, fourth 25 95,8 7+/-0,63 are not replaced in 114+/-0,002 west ^*0,080+/-0,009 ^*

50 95,8 3+/-0,38 ^*0,083+/-0,006 ^{* *}P＜＜0,05; ^*P＜＜0,01 (U de Mann Whitney check).

Therefrom as can be seen, when giving M.H.A.A.B.W (25,50 and 100mg/kg), the gastric juice of Pilorous ligation Mus obviously reduces, and lowest dose level 125mg/kg has obtained maximum efficiency.25 and the west of 50mg/kg obviously reduce the gastric juice volume and also lower H for fourth ⁺Ion concentration.

This test explanation M.H.A.A.B.W (25,50 and 100mg/kg) obviously suppresses the gastric juice volume in the Pilorus model, and does not influence H ⁺Ion concentration, in fact this overcome the western inhibition H that replaces fourth or omeprazol class ⁺The ion secretion effect.Embodiment 9 has used sweet inductive pleuritis of chondrus ocellatus Holmes and cotton grain model in order to confirm the antiinflammatory action of M.H.A.A.B.W, and this is a model of estimating anti-inflammatory drug.

Cotton grain: the above same procedure of testing of the male Sprague Dauley Mus of heavy 200-250g is adapted to laboratory condition.Orally give the suspension of M.H.A.A.B.W in 2%Tween 20/ water, and indometacin is dissolved in 5% sodium bicarbonate.The two all passes through the administration of stomach feeding tube in 6 days after implanting Cotton Gossypii.Mus is divided into 5 test group randomly: 1) contrast (carrier); 2,3 and 4) give 25,50 and the M.H.A.A.B.W of 100mg/kg, 5 respectively) give the indometacin of 3mg/kg.Mus is used etherization earlier, does a kerf at the back.The Cotton Gossypii sheet of under local anaesthesia 50mg being sterilized is implanted subcutaneous, last sew up wound.

After the administration in the end a day, anesthetized animal in the ether atmosphere is carefully peeled off cotton grain, and drying is 24 hours in 60 ℃ of stoves.From cotton grain is heavy, deduct cotton sheet heavy (50mg).Compare with the cotton grain that forms in the matched group, calculate and suppress percentage rate.M.H.A.A.B.W is shown in Table 13 the exercising result of cotton grain.Table 13

Take the effect of M.H.A.A.B.W to cotton grain model

The heavy suppression ratio of the cotton grain of therapeutic dose n

(mg/kg) (mg) (％)

Contrast 8 258+0.12

M.H.A.A.B.KY 25 8 138＋0.05 ^* 46.51

50 7 162＋0.11 ^** 54.7

100 9 117＋0.04 ^** 54.7

Indometacin 35 79+0.02 ^*69.3

^*P＜0.05, ^*P＜0.01 (U de Mann whithey check)

The inductive pleuritis of carragenines: use male Sprague Dawley rat (200,250g), make it adapt to laboratory condition 15 days with water that adds Libitum and food.Test fasting in preceding 12 hours.Give the suspension of M.H.A.A.B.W in 10mg/ml Radix Acaciae senegalis/water, and 30 carbon alcohol, and indometacin is dissolved in 5% sodium bicarbonate.

All administrations were all carried out with the stomach feeding tube before the injection carragenines in 1 hour.Mus is divided into following test group at random: 1) matched group (carrier); 2), 3) and 4) give 25,50 and the M.H.A.A.B.W of 100mg/kg respectively; 5) indometacin of 10mg/kg 30 carbon pure and mild 6 of 50mg/kg).

In order to induce edema, use the etherization rat, at the solution of intrapleural injection 0.3ml carragenines in 1% saline.After 5 hours, put to death animal, collect transudate, survey its volume.Calculate the edema suppression ratio with following formula:

Suppression ratio (%)=1-(VT) is (VC) * 100 wherein:

The edema volume of VT=treatment treated animal

The result of this test of edema volume of VC=control animals represents in 14.

Table 14

The effect of oral M.H.A. A.B.W in the sweet pleuritis that brings out of chondrus ocellatus Holmes

Therapeutic dose n exudate volume suppression ratio

(mg/kg) (ml) (％)

Contrast 15 1.24+0.30

M.H.A.A.B.W 25 10 1.13＋0.39 8.9

50 10 0.98＋0.36 20.9

100 11 0.85＋0.28 ^* 31.4

30 carbon alcohol, 100 10 0.96+0.30 ^*27.4

Indometacin 10 14 0.50+0.15 ^{* *}59.6 ^*P＜0.05 ^{* *}P＜0.001 (U de Mann Whitney check).

From then on as can be seen, under maximum test dose, M.H.A.A.B.W shows medium antiinflammatory action in showing; to the pleural exudate suppression ratio is 23.3%; this is observed maximum effect, and indometacin (10mg/kg) produces protectiveness usefulness to edema, and suppression ratio is 59.6%.30 carbon alcohol are also effective, but its M.H.A.A.B.W that renders a service this similar dosage is low.

These results show that M.H.A.A.B.W ratio in cotton grain model is more effective in the inductive pleuritis model of carragenines.The distinctive points of these two tests is: most important feature is cell migration (neutrophil) in the cotton grain model, and the pleuritis model that carragenines brings out is that inflammatory exudate (increases the blood vessel penetrance, cause edema), back one effect is closely related with prostaglandin (E and F).Strong chemotactic (chemiotactic) effect of inflammatory mediator leukotrienes, particularly LTB4 performance polymorphonuclear leukocyte produces gathering, depolymerization and the lisosomal enzyme that dissociates from them.

A kind of possible explanation of this fact is the synthetic or release that M.H.A.A.B.W suppresses Leukotrienes to a certain extent, produces antiinflammatory action (inhibition cell migration), but also makes amino acid metabolism form elementary prostaglandin, and its mediation transudate increases.This means that M.H.A.A.B.W is only effective in relating to the process of cell migration as antiinflammatory.Embodiment 10

The blended one group of Cavia porcellus albines of the both sexes of average weight 280-350g freely drinks water and gets food, adapts to one week of laboratory condition.Intraperitoneal is given sodium phenobarbital (50mg/kg) with these Animal Anesthesia.One conduit is inserted in the trachea, link (55 breaths/min) with respiratory pump.Volume with 3-4ml is manually ventilated to animal.Measure tracheal pressure with the pressure transmitter that is connected with monitor (MP 0.5).

Giving PAF (40 μ g/Kg) preceding, earlier with M.H.A.A.B.W of the present invention by the intravenous route administration, dosage is 10,1 and 0.5mg/kg.In order to measure the effect of M.H.A.A.B.W to the inductive histamine response of PAF, in the M.H.A.A.B.W administration preceding 10 minutes, give animal PAF or saline (contrast), just produced after a while and repeatably organized ammonia react (2-12 μ g/kg.i.v).Gained the results are shown in table 15 and 16.

Table 15

M.H.A.A.H.W is to the effect of the inductive bronchoconstriction of PAF in the Cavia porcellus

The maximum bronchoconstriction of dosage

(mg/kg) (％)

Contrast-39.60

M.H.A.A.B.W 1.0 13.80 ^*

Contrast-21.28

M.H.A.A.B.W 0.5 20.26?n.s

^*P＜0.05 (U de Mann Whitney check). ^*P＜0.05 (U de Mann Whitney check).

Closed trachea when each off-test has obtained the largest percentage of bronchoconstriction.

Table 16

M.H.A.A.B.W is inductive to PAF in the Cavia porcellus (40ng/kg)

The effect of histamine hiper-reaction

Dosage n bronchoconstriction (%)

(mg/kg) behind the preceding PAF of PAF

Contrast-8 100 157+/-28

M.H.A.A.B.W 10 6 100 105+/-13 ^**

Contrast-5 100 199+/-47.6

M.H.A.A.B.W 1.0 5 100 117+/-25.3 ^**

Contrast-4 100 183+/-18.0

M.H.A.A.B.W 0.5 4 100 172g+/-15n.s.

^*P＜0.01 (U de Mann Whitney check)

In each animal, the histamine response before giving PAF is as 100%.M.H.A.A.B.W impels histamine bronchoconstriction reaction, but obviously lowers the inductive bronchoconstriction of PAF and this reagent in the inductive histamine hiper reaction of 1-10mg/kg concentration.

This M.H.A.A.B.W performance is to the effect of the receptor of PAF and/or Leakotrienes agonist and/or lipoxidase inhibitor.This effect can be explained the antiulcer and the antiinflammatory action of front report.As can be seen, new M.H.A.A.B.W of the present invention does not promptly block the bronchoconstriction effect that does not yet increase group ammonia, and Here it is, and it has overcome anti-group of ammonia effect.Embodiment 11

Character from mellisic senior fat primary alcohol natural mixture of the present invention and comparative study have been carried out from the character (EPO 0,488 928) (being called M.H.A.A.S.C.W) of the mellisic senior fat primary alcohol natural mixture of Caulis Sacchari sinensis at this.This studies have shown that this two mixture are not only different on the relative composition of the number of alcohol and identical alcohol, and its pharmacology performance of test model that is used for pharmacological screening in different tradition is different.The test carried out of reason is as follows for this reason:

A) antiinflammatory action: in order to verify the antiinflammatory action of two mixture, used sweet inductive pleuritis of chondrus ocellatus Holmes and cotton grain model, dosage is respectively 100 and 200mg/kg.

B) antiulcer action: in order to verify the antiulcer action of two mixture, use the test method of describing in the embodiment of the invention 8, surveying mixture reagent is the 25mg/kg body weight.

C) low fat effect: use male new zealand rabbit, be divided into following a) contrast, b) M.H.A.A.B.W (5mg/kg), c) M.H.A.A.S.C.W (5mg/kg), oral month respectively organized.Adopted one time blood sample in per 15 days, to measure lipid parameter (T-CHOL, triglyceride, HDL-C, LDL-C and VLDL-C).

D) ischemia effect:, use the cerebral ischaemia model that produces by the mongolian gerbils artery ligation in order to analyze of the effect of two kinds of mixture to cerebral ischaemia.With the female mongolian gerbils of heavy 60-80g, freely get food and water and make the adaptation laboratory condition.Two kinds of mixture are all by the intraperitoneal administration.With 2%Tween20/ water as carrier.Animal is divided into following test group: 1) contrast (carrier 2%Tween20/ water), 2) M.H.A.A.B.W (200mg/kg) and 3) M.H.A.A.S.C.W (200mg/kg).

With ether atmosphere anesthetized animal, ligation left neck artery.Observed animal 24 hours, the clinical cellulitis shape performance of record brain injury, as turn-take, roll and twitch, and the mortality in process of the test.

E) anti-platelet aggregation effect:, use several the male Sprague Dawley rats of heavy 250-350g in order to verify two mixture to platelet aggregation in the Mus that induces by ADP or collagen.Every kind of mixture was the suspension in Radix Acaciae senegalis/water carrier (1ml/100g body weight) by 4 weeks of stomach feeding tube oral administration.Animal is divided into 3 test group randomly, a) contrast (only using carrier), b) M.H.A.A.B.W (25mg/kg), C) M.H.A.A.S.C.W (25mg/kg).

For measuring the platelet aggregation effect, Mus is anaesthetized with ether.After opening abdominal part, from caval vein, get 5ml blood, mix with 3.8% sodium citrate (9 volume blood with the sodium citrate of 1 volume).Centrifugal blood obtains platelet rich plasma (PRP), and PRP sample part obtained platelet poor plasma (PPP) in centrifugal 15 minutes under 330g.With ADP or collagen-induced platelet aggregation, with Payton aggregometer record.

F) anti-thrombosis function: acted on various blood posts and formed the scale-model investigation anti-thrombosis function.The administration situation is as follows: 1) contrast, 2,3,4) M.H.A.A.B.W (25,50 and 100mg/kg, 5,6,7) M.H.A.A.S.C.W is as (25,50 and 100mg/kg).

The intraperitoneal sodium phenobarbital is with rat fiber crops alcohol.Penetrate hypotonic saline (0.22%NaCl) by the abdomen vein subsequently.After one minute, open abdominal part, expose caval vein, separate and with silk thread with this vein ligation.Sew up abdominal part, open the abdominal cavity after 10 minutes again, in the ligation again of 2cm place, the ligation place downstream first time.Remove immediately then, and vertically cut, removal of thromboses places a wet stove under the room temperature, weigh after one hour.

All pharmacological testing gained results are comprehensively in following table 17.

Table 17: from the natural mixture of peak wax with from the effect comparative test M.H.A.A.S.C.W M.H.A.A.B.W anti-inflammatory of the mixture of sugarcane wax+-the low fat effect-+++anti-ischemia++++antiulcer ++++ anti-platelet aggregation-+antithrombotic forms-+

Active: (+) as seen

(++) is medium

(+++) height

(-) do not have any activity

From these results as can be seen, the pharmacological properties difference of two kinds of senior fat primary alcohol mixture, two kinds of mixture all have antiulcer action, but as can be seen from the table, recently the mixture (M.H.A.A.S.C.W) from sugarcane wax is many effectively from mellisic alcohol mixture (M.H.A.A.B.W).

Claims (14)

1. treatment gastric duodenal ulcer and as the pharmaceutical composition of antiinflammatory, it contains C _24-34The mixture of senior fat primary alcohol as active component, alcohol mixture generally composed as follows:

1-tetracosa carbon alcohol 9.0-15.0%

1-two ten six carbon alcohol 12.0-18.0%

1-octacosanol 13.0-20.0%

1-three ten carbon alcohol 20.0-30.0%

1-three ten two carbon alcohol 13.0-21.0%

1-three ten four carbon alcohols 1.5-3.5%

2. according to the pharmaceutical composition of claim 1, wherein alcohol mixture is more specifically composed as follows:

1-tetracosa carbon alcohol 12.5+/-1.0%

1-two ten six carbon alcohol 14.5+/-1.2%

1-octacosanol 16.5+/-2.0%

1-three ten carbon alcohol 24.6+/-1.6%

1-three ten two carbon alcohol 16.7+/-1.4%

1-three ten four carbon alcohols 2.3+/-0.5%

3. the preparation method from the mellisic natural mixture of forming by high-molecular weight primary aliphatic alcohols of claim 1 or 2 definition, it is characterized in that carrying out the homogeneous phase saponification with alkali metal or alkaline earth metal hydroxide solution after the Cera Flava fusing, and the natural mixture of extraction alcohol, extraction is preferably carried out with following organic solvent in the liquid-solid extraction system: C _6-9Hydrocarbon, C _3-8Ketone, C _1-5Alcohol, halogenated hydrocarbons and aromatic, and their mixture; Alcohol mixture is recrystallization in above-mentioned solvent or its mixture randomly.

4. according to the method for claim 3, it is characterized in that the Cera Flava fusing point is 90-150 ℃, hydroxide concentration is 5-30%, saponification time at least 30 minutes, and the extraction time is 1-20 hours.

5. according to the method for claim 3 or 4, it is characterized in that being used for the saponified hydroxide of Cera Flava is sodium hydroxide, calcium hydroxide or potassium hydroxide.

6. according to the method for claim 3 or 4, it is characterized in that the used hydrocarbon of extraction step is pentane, hexane, heptane or octane.

7. according to the method for claim 3 or 4, it is characterized in that the ketone that is used to extract is acetone, methyl ethyl ketone, methyl butyl ketone or 3-heptanone.

8. according to the method for claim 3 or 4, it is characterized in that the alcohol that is used to extract is methanol, ethanol, normal propyl alcohol, 2-propanol, n-butyl alcohol, 2-butanols, n-amyl alcohol or the tert-butyl alcohol.

9. according to the method for claim 3 or 4, it is characterized in that the aromatic solvent that is used to extract is benzene, toluene, ethylbenzene, phenol or xylol.

10. according to the method for claim 3 or 4, it is characterized in that the halogenated hydrocarbons that is used to extract is dichloromethane, 1,2-dichloroethanes, chloroform, trichloroethane, 1,2-dichloropropane or 1,2,3-trichloropropane.

11. medicament preparation such as tablet, capsule or gragees, it is characterized in that they contain 0.5～25.0% (weight) claim 1 or 2 from mellisic high molecular primary aliphatic alcohols natural mixture as active component, also contain filler, agglutinant, disintegrating agent, lubricant and other medicines auxiliary shape agent.

12. the purposes of the medicament preparation treatment gastric duodenal ulcer that derives from mellisic senior fat primary alcohol natural mixture or claim 11 of claim 1 or 2.

13. power requires the effect as antiinflammatory of 1 or 2 the medicament preparation that derives from mellisic senior fat primary alcohol natural mixture or claim 11.

14. the purposes of the medicament preparation that derives from mellisic senior fat primary alcohol natural mixture or claim 11 of claim 1 or 2, the oral or parenterai administration of natural alcohol mixture wherein, daily dose is 1-100mg, particularly 10-20mg.