CN1180785C - Application of fur seal oil in preparation of medicine for treating diabetes - Google Patents
Application of fur seal oil in preparation of medicine for treating diabetes Download PDFInfo
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- CN1180785C CN1180785C CNB011355212A CN01135521A CN1180785C CN 1180785 C CN1180785 C CN 1180785C CN B011355212 A CNB011355212 A CN B011355212A CN 01135521 A CN01135521 A CN 01135521A CN 1180785 C CN1180785 C CN 1180785C
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- phocae vitulinae
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Abstract
The present invention relates to an application of seal's fat for preparing medicine for treating diabetes. Researchers find that when the seal's fat of the present invention is continuously poured and administrated to stomachs under the dose of 16 to 32 ml/kg, the increased blood sugar can be obviously lowered to diabetes caused by alloxan of mice, and the body weight of mice is obviously increased. Therefore, under the situation that the seal's fat is used as medicine, the seal's fat has obvious effect on treating diabetes at a larger dose.
Description
Technical field
What the present invention relates to is that specifically, what the present invention relates to is the application of Adeps Phocae vitulinae being controlled mammal and people's Rezulin as preparation with the application of Adeps Phocae vitulinae as the preparation antidiabetic drug.
Background technology
As everyone knows, patient's quantity of diseases such as diabetes, hepatopathy, prostate, hyperlipidemia is very huge, and in the practical application, has special positive effect and the medicine of almost being free from side effects is difficult to confirm.
It has been generally acknowledged that patient's quantity that Eskimos and Japanese coastal fisherman suffer from this type of disease is significantly lower than general crowd, research worker believe this phenomenon and eat the fur seal meat that is rich in unsaturated fatty acid with them, oil is relevant.
But, do not have concrete zoopery or clinical effectiveness to be proved so far.
Described in the present invention fur seal typically refers to a kind of mammal of life Newfoundland, North Atlantic Region, the arctic, in their body fats unsaturated fatty acid content up to 25%, and its molecular structure and the intravital fatty acid molecule structural similarity of people.
Adeps Phocae vitulinae of the present invention (claiming seal oil again) is meant that the subcutaneous fat by above-mentioned fur seal refines the all-natural product that forms.
Result of study shows, containing in the body of a large amount of needed by human in the Adeps Phocae vitulinae can not synthetic voluntarily ω-3 be long carbochain polyene unsaturated fatty acid and a spot of Squalene (SPUALENE).
Summary of the invention
The object of the present invention is to provide a kind of with the application of Adeps Phocae vitulinae as the preparation antidiabetic drug.
The Adeps Phocae vitulinae that pharmacological experiment of the present invention adopts is to extract the little yellow oily liquid that forms in the fur seal body, provide in soft capsule (0.5g/ grain) mode by Canadian Atlantic Ocean wholesome food company limited, lot number is 11656, and described Adeps Phocae vitulinae adopts room temperature preservation at laboratory.
For the further composition and the content of clear and definite above-mentioned Adeps Phocae vitulinae, below the composition of Adeps Phocae vitulinae used in the present invention is analyzed and confirmed.
Prior art shows that the ω that extracts-3 is that the chemical main composition of polyene unsaturated fatty acid is EPA, DHA and DPA from Adeps Phocae vitulinae, wherein:
EPA is meant eicosapentaenoic acid;
DPA is meant clupanodonic acid;
DHA is meant docosahexenoic acid;
Research worker of the present invention is carried out the mensuration of projects such as ultra-violet absorption spectrum, infrared absorption spectroscopy, gas chromatogram, gas chromatography-mass spectrometry chromatogram to the Adeps Phocae vitulinae of above-mentioned acquisition, and the chemical constitution of described sample is confirmed, and is specific as follows:
The Adeps Phocae vitulinae test result of samples that the present invention adopts is:
(1) ultra-violet absorption spectrum
With chloroform (analytical pure) sample is made into and carries out the long UV scanning of all-wave behind the 5.9mg/ml solution, the model of employed instrument is U-3210spectrophotometer (Japan manufacturing); Referring to accompanying drawing 1, its result shows that sample has absorption maximum at the 249.8nm place, can prove and contain a large amount of unsaturated bonds in the sample;
(2) infrared absorption spectroscopy
Referring to accompanying drawing 2, its result shows, at 3010cm
-1And 1745cm
-1The strong absorption at place be respectively unsaturated=C-H and＞characteristic absorption of C=O stretching vibration; 1655cm
-1The weak absorption at place meets the stretching vibration characteristic absorption (1665-1635cm of cis-double bonds (R1-C=C-R2)
-1, trans is 1675-1665cm
-1),
722cm
-1The absworption peak at place meets the characteristic absorption (730-665cm of cis-double bonds=C-H bending vibration
-1, trans is 980-960cm
-1).
Comprehensive infrared and ultraviolet spectral analysis can be thought to be mainly the cis non-conjugated fatty acid in the sample.
(3) gas chromatogram
In this detection, the accepted standard product are provided by SIGMA company, are respectively:
Cis-5,8,11,14,17-EICOSAPENTAENOIC ACID METHYL ESTER, methyl eicosapentaenoic acid;
Cis-4,7,10,13,16,19-DOCOSAHEXAENOIC ACID ETHYL ESTER, docosahexaenoic acid ethyl;
Cis-7,10,13,16,19-DOCOSAPENTAENOIC ACID METHYL ESTER, clupanodonic acid methyl ester;
Sample is the Adeps Phocae vitulinae of above-mentioned acquisition, and analyzing fatty acid with the GC method need specifically be to get described sample 80 μ l with its ethyl esterization in order to gasification, add the NaOH-ethanol liquid 1ml of 0.5ml/l, fill N2, jump a queue, jolting adds boron trifluoride ethanol liquid 1.5ml to little oil droplet complete obiteration in 50 ℃ of water-baths.Mix homogeneously is put 5min in 50 ℃ of water-baths, take out cooling, and adding normal heptane 1ml, saturated NaCl is the 2ml mix homogeneously, layering.Get upper strata liquid in another test tube, add a small amount of anhydrous NA2SO4, fill N2, treat that in 4 ℃ of placements GC analyzes.
Employed instrument is HP6890;
GC conditions: glass filled post 2m * 3mm, 10%DEGS pickling, 101 white carriers, carrier gas N2, flow velocity 25ml/min, detector FID, 180 ℃ of column temperatures, 210 ℃ of vaporizers, 270 ℃ of detectors, sample size 1ul.
With standard substance normal hexane standardize solution, be configured to 2.5mg/ml solution, precision is measured 1ul and is carried out the gas phase assay, and same method is measured sample, and the result is that the retention time of sample and reference substance is consistent.
Can be referring to accompanying drawing 3-7.
(4) gas-matter coupling method
The instrument that adopts is a U.S. Trace 2000GC/Trace MS type gas chromatograph-mass spectrometer, and all reagent are analytical pure.
Experiment condition is:
GC conditions: the PEG-20M glass capillary column (2m * 0.32mm); Column temperature keeps 1min for 40 ℃, is warming up to 200 ℃ with 10 ℃/min, keeps 5min, is warming up to 210 ℃ with 30 ℃/min again, keeps 2min; Carrier gas N2 presses 3.0kpa before the post, split ratio 1: 30, injector temperature ℃ 250 ℃; 250 ℃ of ion source temperatures, ionization voltage 70Ev, mass scanning scope 350AmU.S-1; Sample size 1ml.
Through mass spectrogram analysis, determine that the molecular weight of sample and standard substance conforms to.
So far, molecular weight, composition, the content and structure of of the Adeps Phocae vitulinae that the present invention is adopted have obtained confirming.
The preparation of material therefor in the pharmacological experiment test of the present invention.
One, Adeps Phocae vitulinae
Describedly in foregoing, describe in detail, press the 25g body weight during experiment and irritate stomach 0.8ml (high dose group use stock solution), or 0.6ml (in, low dosage uses dilute liquid medicine).
Two, positive control drug
The positive control drug that uses among the present invention is phenformin hydrochloride tablet (JIANGZHILING PIAN), every content 25mg, and lot number 990223, the accurate word (1995) of medicine is defended No. 034024 in the authentication code Shandong, is produced by Shandong medicine system factory, purchases in pharmacy of Beijing synergism pharmaceutical factory.Before the experiment tablet is crushed in mortar, add distilled water while grinding, make the suspendible medicinal liquid of 5mg/ml, the mouse stomach volume is pressed the 0.5ml/g body weight.
Three, model drug
The model drug of using among the present invention is alloxan.Acne, lot number 39H1482 gives birth to preface by SIGMA company, is bought on behalf by supply and marketing section of this institute.4 ℃ of preservations take by weighing alloxan before the experiment on 1/0,000 balances, make the suspendible medicinal liquid of 7mg/ml, preserve in the ice bath, and the mouse tail vein injection volume is pressed the 0.2ml/20g body weight.
Four, blank group mice
Every of the blank group mice of using among the present invention is irritated stomach 0.5ml distilled water.
Five, biochemical reagents and other reagent
The biochemical reagents that use among the present invention and other pack are drawn together:
The glucose assays test kit is the production of the smart coin chemicals of Beijing northization company limited;
Normal saline, olive oil are purchased in Beijing pharmacy.
Six, experimental animal
Experimental animal among the present invention is a cleaning level Kunming mouse, mice volume 24-26g, and the animal quality quality certification number: the capital is moving is betrothed to (2000) No. 012, is provided by the Nat'l Pharmaceutical ﹠ Biological Products Control Institute experimental center.Observed two days before the experiment, choosing is healthy, and active animal is used for test.
In order to confirm that Adeps Phocae vitulinae of the present invention is in the curative effect aspect the treatment diabetes, carried out following animal experiment, experiment purpose is to understand described Adeps Phocae vitulinae alloxan is caused the pharmacotoxicological effect and the effect of mice diabetes model, and concrete method and result are as follows:
Get 120 of mices, body weight 24-26g, every mice is by tail vein injection alloxan 70mg/kg, divide cage at random, every mice all makes marks, after 72 hours (the mice fasting is 4 hours before the blood sampling), venous plexus is got blood 0.4-0.5ml at the bottom of the eye socket, it is centrifugal to leave standstill the back, gets serum glucose assays kit measurement blood glucose, and the mensuration wavelength is 550nm.
Choose blood glucose value 60 of the mices of 250-600mg/ml, be divided into 5 groups, make the difference of every group of mean blood glucose be not more than 20mg/dL.Tangible polyuria symptom appears in mice.
Enforcement is divided into 6 groups: i.e. blank group, alloxan model 70mg/kg group, positive control drug JIANGZHILING PIAN 100mg/kg group, Adeps Phocae vitulinae 32ml/kg group, 16ml/kg, 8ml/kg group, 12 every group.
Blank group and model control group are only irritated stomach feedwater 0.5ml/ every day, three dosage groups of Adeps Phocae vitulinae and positive control drug group gastric infusion every day 1 time, and successive administration 14 days is not seen drug-induced untoward reaction during the administration.Full 14 days of administration, carry out the detection of following index:
1, the mensuration of blood glucose: mice fasting 4 hours before time administration, 1.5 hours all mices venous plexus at the bottom of the eye socket is got blood 0.4-0.5ml after the administration, and it is centrifugal to leave standstill the back, gets serum and measures kit measurement blood glucose with Fructus Vitis viniferae, and the mensuration wavelength is 550nm.
Table 1 Adeps Phocae vitulinae is to the influence () of blood glucose
| Group | Dosage ml/kg | Number of animals n | ?X+SD.mg/dL | The P value | |
| Blank | — | 10 | ?136.21±19.29 | ||
| The | 70mg | 10 | ?554.0±101.79 * | ---- | |
| JIANGZHILING PIAN | | 10 | ?378.9±208.89 | ＜0.05 | |
| Adeps Phocae vitulinae | 8 16 32 | 10 10 10 | ?589.56±85.6 ?383.3±153.82 ?333.9±127.56 | ＞0.05 ＜0.05 ＜0.01 |
Compare with the blank group
*P＜0.01
Adeps Phocae vitulinae is to the influence (two) of blood glucose
| Group | Dosage ml/kg | Number of animals n | ?X+SD.mg/dL | The P value |
| Blank | — | ?10 | ?104.15±3.44 | |
| The model contrast | 70mg | ?10 | ?529.57±158.10 * | ?---- |
| JIANGZHILING PIAN | 100mg | ?10 | ?380.13±113.75 | ?P＜0.05 |
| | 8 16 32 | ?10 ?10 ?10 | ?528.58±127.31 ?490.02±89.93 ?385.32±113.75 | ?P＞0.05 ?P＞0.05 ?P＜0.05 |
Compare with the blank group
*P＜0.01
The above results shows that the blood sugar content of blank group is in range of normal value; The numerical value of model control group raises then very obvious; And positive controls and Adeps Phocae vitulinae 32ml/kg, the numerical value of 16ml/kg group all has tangible reduction, has significant differences and significant difference (p＜0.01, p＜0.05) with the analysis of model control group comparative statistics.
2, the mensuration of body weight
Before inferior administration, take by weighing and respectively organize the mice body weight, measure the significance of respectively organizing body weight meeting content difference with two groups of t checks.The result sees table 2 for details.
Table 2 Adeps Phocae vitulinae is to the influence of body weight
| Group | Dosage ml/kg | Number of animals n | ?X+SD.mg/dL | The P value |
| Blank | — | ?10 | ?32.3±1.70 | |
| The model contrast | 70mg | ?10 | ?20.5±4.01 * | ?---- |
| JIANGZHILING PIAN | 100mg | ?10 | ?27.18±3.32 | ?P＜0.01 |
| | 8 16 32 | ?10 ?10 ?10 | ?22±5.00 ?26±2.10 ?27.2±1.99 | ?P＞0.05 ?P＜0.01 ?P＜0.01 |
Compare with the blank group
*P＜0.01
The above results shows that the body weight of blank group is in range of normal value; The numerical value of model control group reduces then very obvious; And positive controls and Adeps Phocae vitulinae 32ml/kg, the numerical value of 16ml/kg group all has obvious reduction, has significant differences and significant difference and significance (P＜0.01) with the analysis of model control group comparative statistics.
From above experiment as can be seen, Adeps Phocae vitulinae of the present invention is under 16-32ml/kg dosage, and continuously gastric infusion is 14 days, and the diabetes to due to the mice alloxan can make the blood glucose of rising significantly reduce; Obviously increase the body weight of mice, confirm that this medicine than under the heavy dose, has the effect of tangible antagonism alloxan induced mice diabetes.
By above test as can be seen, utilize alloxan to cause the model of mice diabetes, to Adeps Phocae vitulinae under 8ml/kg-32ml/kg dosage, 14 days pharmacodynamic action of continuous irrigation stomach is observed, found that Adeps Phocae vitulinae of the present invention has blood glucose that obvious reduction raises and effect that can weight increase numerical value during diabetes in treatment, shows that Adeps Phocae vitulinae of the present invention has the effect of tangible antagonism treatment of diabetes.
Description of drawings
Below be description of drawings of the present invention,, can more be expressly understood the present invention by description of drawings and in conjunction with above-mentioned specific descriptions, specific as follows:
Accompanying drawing 1 is the uv absorption spectrogram of Adeps Phocae vitulinae of the present invention;
Accompanying drawing 2 is infrared absorption spectras of Adeps Phocae vitulinae of the present invention;
Accompanying drawing 3-5 is the gas chromatogram of the standard substance of the EPA methyl ester that uses among the present invention, DHA ethyl ester, DPA methyl ester;
Accompanying drawing 6-7 is the gas chromatogram of Adeps Phocae vitulinae of the present invention;
Claims (3)
1. the Main Ingredients and Appearance Adeps Phocae vitulinae that is eicosapentaenoic acid, clupanodonic acid and docosahexenoic acid is in the application of preparation antidiabetic drug.
2. application according to claim 1 is characterized in that the dosage of the described 8-32ml/kg of being applied as.
3. application according to claim 1 and 2 is characterized in that the dosage of the described 16-32ml/kg of being applied as.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011355212A CN1180785C (en) | 2001-09-30 | 2001-09-30 | Application of fur seal oil in preparation of medicine for treating diabetes |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB011355212A CN1180785C (en) | 2001-09-30 | 2001-09-30 | Application of fur seal oil in preparation of medicine for treating diabetes |
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| Publication Number | Publication Date |
|---|---|
| CN1410077A CN1410077A (en) | 2003-04-16 |
| CN1180785C true CN1180785C (en) | 2004-12-22 |
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| CNB011355212A Expired - Fee Related CN1180785C (en) | 2001-09-30 | 2001-09-30 | Application of fur seal oil in preparation of medicine for treating diabetes |
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| CN104814950A (en) * | 2010-11-09 | 2015-08-05 | 持田制药株式会社 | Agent for inhibiting elevation in postprandial blood glucose level |
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Owner name: CHINA BIOCHEMICAL PHARMACEUTICAL INDUSTRY ASSOCIA Free format text: FORMER OWNER: CHINESE VERIFICATION OFFICE OF MEDICINE AND BIOLOGICAL PRODUCTS Effective date: 20070803 |
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Effective date of registration: 20070803 Address after: 100801. Room 45, No. 714, main street, Fuxing gate, Beijing Patentee after: China Biochemical Pharmaceutical Industry Association Address before: 100061 No. 2, West Lane, Tiantan, Beijing Patentee before: National Institute for the Control of Pharmaceutical and Biological Products |
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Granted publication date: 20041222 Termination date: 20130930 |