CN105726624A - Pharmaceutical composition for treating diabetes - Google Patents
Pharmaceutical composition for treating diabetes Download PDFInfo
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- CN105726624A CN105726624A CN201610285707.0A CN201610285707A CN105726624A CN 105726624 A CN105726624 A CN 105726624A CN 201610285707 A CN201610285707 A CN 201610285707A CN 105726624 A CN105726624 A CN 105726624A
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- ethanol
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- saponin
- polysaccharide
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- XZWYZXLIPXDOLR-UHFFFAOYSA-N metformin hydrochloride Natural products CN(C)C(=N)NC(N)=N XZWYZXLIPXDOLR-UHFFFAOYSA-N 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
- A61K36/481—Astragalus (milkvetch)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation or decoction
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
- A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/30—Extraction of the material
- A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K2236/50—Methods involving additional extraction steps
- A61K2236/55—Liquid-liquid separation; Phase separation
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Medical Informatics (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention relates to a pharmaceutical composition for treating diabetes, and belongs to the technical field of medicines. The pharmaceutical composition is prepared from the following medicines in parts by weight: 30-70 parts of 60%-70% polysaccharide, 25-60 parts of 70%-85% saponin and 5-10 parts of 55%-70% flavones. Common fenugreek seeds are adopted as a main raw material; and the astragalus membranaceus is taken as an auxiliary material. The invention provides a method for effectively separating and purifying the polysaccharide, the saponin and the flavones in the common fenugreek seeds and the astragalus membranaceus. Through intensive study and combination of the polysaccharide, the saponin and the flavones, the effective component content is high; the toxicity and the side effect of the medicine are removed; the quality is easy to control; the blood glucose of diabetic animals can be significantly reduced; and the pharmaceutical composition has good prevention and treatment effects on diabetes and complications. The diabetes is treated by combined components of the traditional Chinese medicines; the preparation technology is easy to industrialize; the pharmaceutical composition has great and profound meaning for modernization of the traditional Chinese medicine and marching for the international market.
Description
Technical field
The invention belongs to pharmaceutical technology field, particularly relate to a kind of traditional Chinese medicine effective ingredient compatibility for preventing and treating diabetes.
Background technology
Diabetes (DM) are defect of insulin secretion or insulin action obstacle causes a kind of be feature with hyperglycemia metabolic disease, along with the metabolism disorder of sugar, fat, protein etc..Long-term hyperglycemia and metabolism disorder and then concurrent body tissue organ lesion, including kidney, cardiovascular, gallbladder, eye, nervous system damage and skin repeated infection etc..DM typical clinical symptom is " three-many-one-little ", i.e. polyuria, polydipsia, polyphagia and lose weight.The change of mode and the increase of operating pressure is drawn together along with raw, diabetics just increases with surprising rapidity, the current whole world has more than the population of 2.85 hundred million and suffers from diabetes, makes a definite diagnosis that to have half in the patient of diabetes be crowd between 20～60 years old, and this numeral will rise violently 4.35 hundred million to the year two thousand thirty.Diabetes have become one of three serious diseases of serious harm human health mutually arranged side by side with cardiovascular and cerebrovascular vessel, tumor, and in worldwide, the trend of diabetes expansion is severe all the more, thus the drug research preventing and treating diabetes is very urgent.
Hypoglycemic drug is many based on chemical synthetic drug in the market, it is easy to produces dependency, and easily produces complication.And blood sugar lowering effective ingredient has following characteristics in Chinese medicine: 1. curative effect is gentle, stable and lasting；2. toxic and side effects is very little；3. the function such as blood sugar lowering and protection renal is generally had concurrently；4. the usual composite use of multiple sugar-lowering components has the effect of Synergistic；5. most of Chinese medicine blood sugar reducing component stable in properties, can pass through oral administration, taking convenience.
In recent years, the research about the Chinese medicine compound clinical observation report for the treatment of diabetes and experimentation and to single medicine and traditional Chinese medicine effective ingredient has become a big focus in diabetes study field.Owing to traditional Chinese medicine effective ingredient compatibility compound treatment can giving consideration to both the incidental and fundamental, energy its complication of effectively preventing while treatment diabetes primary symptom, and mechanism of action and approach have polytropism, multi-level pharmacology, being not likely to produce the situations such as insulin resistant, the advantage of traditional Chinese medicine effective ingredient compatibility compound treatment diabetes and complication day by day manifests.Know that the Chinese medicine having definite curative effect mainly has Semen Trigonellae, the Radix Astragali, Radix Ginseng, Radix Salviae Miltiorrhizae, Radix Notoginseng, the Radix Rehmanniae, Radix Rehmanniae Preparata, Radix Et Rhizoma Rhei, Herb Gynostemmae Pentaphylli etc. after deliberation at present.
Semen Trigonellae is the dry mature seed of leguminous plant Semen Trigonellae, in Asia, Mediterranean and African country is widely cultivated and for food, field of health care products, the report that Semen Trigonellae has hypoglycemic activity is long-standing, and its extract and germination goods all have the effect of well treatment diabetes and hyperlipidemia.The hypoglycemic main composition of Semen Trigonellae includes fenugreek polysaccharide, saponin, flavone.Wherein Fenugreek Polysaccharides can effectively control blood glucose, and it is by suppressing sugar digestive enzyme activity in intestinal, delay sugar absorption, promote hepatic glycogen synthesis, improving the approach blood sugar lowering such as insulin resistant.Trigoneoside is by repairing impaired islets of langerhans, suppress the digested enzymatic activity of intestinal, increase hepatic glycogen content, suppressing the approach blood sugar lowering such as glyconeogenesis enzymatic activity.Diabetic mice pancreas ponderal index can be improved, promote the approach blood sugar lowering such as synthesis of hepatic glycogen.And flavone component is by repairing impaired islets of langerhans, promote insulin secretion, increase hepatic glycogen content, improving the approach performance hypoglycemic activities such as insulin resistant.The Radix Astragali is traditional traditional tonic medicine, and modern pharmacological research display astragalus polysaccharides, saponin, flavonoid component have blood sugar lowering, improve carbohydrate metabolism, insulin resistant and inhibited oxidation glycosylation, have significantly protection Diabetic Nephropathy function simultaneously.
Summary of the invention
The present invention provides a kind of pharmaceutical composition treating diabetes, is used for treating diabetes.
The present invention adopts the technical scheme that: be prepared by what the medicine of parts by weight was made:
60%～70% polysaccharide 30～70 parts；
70%～85% saponin 25～60 parts；
55%～70% flavone 5～10 parts.
60%～70% polysaccharide of the present invention is prepared by what method prepared:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder by the weight ratio powder of 10:1～5, adds 8～12 times amount 60-90 DEG C of petroleum ether, ultrasonic degreasing 40～60min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) Polyose extraction: weigh defat medical material, add 5～20 times amount 60%～80% ethanol, soak 0.5～1h, circumfluence distillation 3 times, each 1～2h, extracting solution is evaporated to without alcohol taste, and after extraction, remaining medicinal residues retain, dry, add 5～15 times amount distilled water, extract 60～120min, 10000r/min centrifugal 10min when 10 DEG C of magnetic agitation, collect supernatant；
(3) polysaccharide purification: 60%～80% ethanol precipitate with ethanol, stands overnight, and 4000r/min is centrifuged 10min, supernatant discarded, and precipitation distilled water fully redissolves, lyophilization, obtains polysaccharide purification component, determination of color polyoses content.
55%～70% flavone of the present invention is prepared by what method prepared:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder by the weight ratio powder of 10:1～5, adds 60 DEG C～90 DEG C petroleum ether of 8～12 times amount, ultrasonic degreasing 40～60min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) extract: weigh defat medical material, add 5～20 times amount 60%～80% ethanol, soak 0.5～1h, circumfluence distillation 3 times, each 1～2h, extracting solution is evaporated to without alcohol taste, with distilled water, concentrated solution is settled to 0.2g crude drug/mL, after extraction, remaining medicinal residues retain, dry, polysaccharide to be extracted；
(3) loading just purification: by step (2) gained 0.2g crude drug/ml extracting solution with 1BV/h flow velocity loading to the polyamide resin column handled well, d=10cm, h=60cm, the extracting liquid volume of 1BV=1.5 times, with 4BV water with 4BV/h flow velocity eluting remove impurity, then with 5BV60%～80% ethanol with 4BV/h flow velocity eluting, eluent is evaporated to without alcohol taste, it is dissolved in water and is settled to the volume identical with first time constant volume, treating that D101 macroporous adsorbent resin is further purified；
(4) loading repurity: by step (3) gained just purification of flavone solution with 1BV/h flow velocity loading to the D101 resin column handled well, d=10cm, h=60cm, the extracting liquid volume of 1BV=1.5 times, with 5BV20%～40% ethanol with 4BV/h flow velocity eluting, starts to collect eluent from 2BV volume, after being evaporated to a small amount of volume, lyophilization, obtains flavone purified components, determination of color flavones content.
70%～85% saponin of the present invention is prepared by what method prepared:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder by the weight ratio powder of 10:1～5, adds 8～12 times amount 60-90 DEG C of petroleum ether, ultrasonic degreasing 40～60min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) extract: weigh defat medical material, add 5～20 times amount 60%～80% ethanol, soak 0.5～1h, circumfluence distillation 3 times, each 1～2h, extracting solution is evaporated to without alcohol taste, with distilled water, concentrated solution is settled to 0.2g crude drug/mL, after extraction, remaining medicinal residues retain, dry, polysaccharide to be extracted；
(3) saponin purification: by step (2) gained 0.2g crude drug/ml extracting solution with 1BV/h flow velocity loading to the DM130 resin column handled well, d=10cm, h=60cm, the extracting liquid volume of 1BV=1.5 times, after absorption 2h, with 3BV water with 4BV/h flow velocity eluting remove impurity, remove impurity is continued with 4BV/h flow velocity with 5BV20%～40% ethanol, again with 5BV60%～80% ethanol with 4BV/h flow velocity eluting saponin, the concentrated lyophilization of eluent, obtain saponin purification fractions, determination of color saponin content.
The medicinal raw material of the present invention is based on Semen Trigonellae, the Radix Astragali is auxiliary, provide one and efficiently separate purification Semen Trigonellae, polysaccharide in the Radix Astragali, saponin, the method of flavonoid component, through further investigation, pass through polysaccharide, saponin, the compatibility of flavonoid component, active constituent content is high, and eliminate the Side effect of medicine, quality is easily controlled, diabetic animal blood glucose can be significantly reduced, diabetes and complication thereof there is good preventive and therapeutic action, the drug matching component treatment diabetes that the present invention adopts, preparation technology is prone to industrialization, for the modernization of Chinese medicine with move towards international market there is important and far-reaching meaning.
Accompanying drawing explanation
Fig. 1 is rat kidney tissue HE coloration result figure；
Fig. 2 is rat blood serum triglyceride determination result figure；
Fig. 3 is rat blood serum T-CHOL measurement result figure；
Fig. 4 is rat blood serum low-density LP determination result figure.
Detailed description of the invention
Embodiment 1
It is prepared by what the medicine of parts by weight was made:
60%～70% polysaccharide 30 parts；
70%～85% saponin 25 parts；
55%～70% flavone 5 parts；
Embodiment 2
It is prepared by what the medicine of parts by weight was made:
60%～70% polysaccharide 50 parts；
70%～85% saponin 42.5 parts；
55%～70% flavone 7.5 parts；
Embodiment 3
It is prepared by what the medicine of parts by weight was made:
60%～70% polysaccharide 70 parts；
70%～85% saponin 60 parts；
55%～70% flavone 10 parts；
Embodiment 4
It is prepared by what the medicine of parts by weight was made:
60%～70% polysaccharide 10 parts；
70%～85% saponin 4 parts；
55%～70% flavone 1 part；
Embodiment 5
It is prepared by what the medicine of parts by weight was made:
60%～70% polysaccharide 12 parts；
70%～85% saponin 8 parts；
55%～70% flavone 1 part；
Embodiment 6
It is prepared by what the medicine of parts by weight was made:
60%～70% polysaccharide 3 parts；
70%～85% saponin 6 parts；
55%～70% flavone 1 part；
In above-described embodiment:
60%～70% described polysaccharide is prepared by what method prepared:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder by the part by weight powder of 10:1, adds 60 DEG C of petroleum ether of 8 times amount, ultrasonic degreasing 40min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) Polyose extraction: weigh defat medical material, add 5 times amount 60% ethanol, soak 0.5h, circumfluence distillation 3 times, each 1h, extracting solution is evaporated to without alcohol taste, and after extraction, remaining medicinal residues retain, dry, add 5 times amount distilled water, extract 60min, 10000r/min centrifugal 10min when 10 DEG C of magnetic agitation, collect supernatant；
(3) polysaccharide purification: 60% ethanol precipitate with ethanol, stands overnight, and 4000r/min is centrifuged 10min, supernatant discarded, and precipitation distilled water fully redissolves, lyophilization, obtains polysaccharide purification component, determination of color polyoses content；
Or:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder by the weight ratio powder of 10:3, adds 75 DEG C of petroleum ether of 10 times amount, ultrasonic degreasing 50min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) Polyose extraction: weigh defat medical material, add 12 times amount 70% ethanol, soak 0.8h, circumfluence distillation 3 times, each 1.5h, extracting solution is evaporated to without alcohol taste, and after extraction, remaining medicinal residues retain, dry, add 10 times amount distilled water, extract 90min, 10000r/min centrifugal 10min when 10 DEG C of magnetic agitation, collect supernatant；
(3) polysaccharide purification: 70% ethanol precipitate with ethanol, stands overnight, and 4000r/min is centrifuged 10min, supernatant discarded, and precipitation distilled water fully redissolves, lyophilization, obtains polysaccharide purification component, determination of color polyoses content.
Or:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder by the weight ratio powder of 10:5, adds 90 DEG C of petroleum ether of 12 times amount, ultrasonic degreasing 60min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) Polyose extraction: weigh defat medical material, add 20 times amount 80% ethanol, soak 1h, circumfluence distillation 3 times, each 2h, extracting solution is evaporated to without alcohol taste, and after extraction, remaining medicinal residues retain, dry, add 15 times amount distilled water, extract 120min, 10000r/min centrifugal 10min when 10 DEG C of magnetic agitation, collect supernatant；
(3) polysaccharide purification: 80% ethanol precipitate with ethanol, stands overnight, and 4000r/min is centrifuged 10min, supernatant discarded, and precipitation distilled water fully redissolves, lyophilization, obtains polysaccharide purification component, determination of color polyoses content.
55%～70% described flavone is prepared by what method prepared:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder by the weight ratio powder of 10:5, adds 60 DEG C of petroleum ether of 8 times amount, ultrasonic degreasing 40min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) extract: weigh defat medical material, add 5 times amount 60% ethanol, soak 0.5h, circumfluence distillation 3 times, each 1h, extracting solution is evaporated to without alcohol taste, with distilled water, concentrated solution is settled to 0.2g crude drug/mL, after extraction, remaining medicinal residues retain, dry, polysaccharide to be extracted；
(3) loading just purification: by step (2) gained 0.2g crude drug/ml extracting solution with 1BV/h flow velocity loading to the polyamide resin column handled well, d=10cm, h=60cm, the extracting liquid volume of 1BV=1.5 times, with 4BV water with 4BV/h flow velocity eluting remove impurity, then with 5BV60% ethanol with 4BV/h flow velocity eluting, eluent is evaporated to without alcohol taste, it is dissolved in water and is settled to the volume identical with first time constant volume, treating that D101 macroporous adsorbent resin is further purified；
(4) loading repurity: by step (3) gained just purification of flavone solution with 1BV/h flow velocity loading to the D101 resin column handled well, d=10cm, h=60cm, the extracting liquid volume of 1BV=1.5 times, with 5BV20% ethanol with 4BV/h flow velocity eluting, starts to collect eluent from 2BV volume, after being evaporated to a small amount of volume, lyophilization, obtains flavone purified components, determination of color flavones content；
Or:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder by the weight ratio powder of 10:3, adds 75 DEG C of petroleum ether of 10 times amount, ultrasonic degreasing 50min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) extract: weigh defat medical material, add 12 times amount 70% ethanol, soak 0.8h, circumfluence distillation 3 times, each 1.5h, extracting solution is evaporated to without alcohol taste, with distilled water, concentrated solution is settled to 0.2g crude drug/mL, after extraction, remaining medicinal residues retain, dry, polysaccharide to be extracted；
(3) loading just purification: by step (2) gained 0.2g crude drug/ml extracting solution with 1BV/h flow velocity loading to the polyamide resin column handled well, d=10cm, h=60cm, the extracting liquid volume of 1BV=1.5 times, with 4BV water with 4BV/h flow velocity eluting remove impurity, then with 5BV70% ethanol with 4BV/h flow velocity eluting, eluent is evaporated to without alcohol taste, it is dissolved in water and is settled to the volume identical with first time constant volume, treating that D101 macroporous adsorbent resin is further purified；
(4) loading repurity: by step (3) gained just purification of flavone solution with 1BV/h flow velocity loading to the D101 resin column handled well, d=10cm, h=60cm, the extracting liquid volume of 1BV=1.5 times, with 5BV30% ethanol with 4BV/h flow velocity eluting, starts to collect eluent from 2BV volume, after being evaporated to a small amount of volume, lyophilization, obtains flavone purified components, determination of color flavones content；
Or:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder by the weight ratio powder of 10:5, adds 90 DEG C of petroleum ether of 12 times amount, ultrasonic degreasing 60min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) extract: weigh defat medical material, add 20 times amount 80% ethanol, soak 1h, circumfluence distillation 3 times, each 2h, extracting solution is evaporated to without alcohol taste, with distilled water, concentrated solution is settled to 0.2g crude drug/mL, after extraction, remaining medicinal residues retain, dry, polysaccharide to be extracted；
(3) loading just purification: by step (2) gained 0.2g crude drug/ml extracting solution with 1BV/h flow velocity loading to the polyamide resin column handled well, d=10cm, h=60cm, the extracting liquid volume of 1BV=1.5 times, with 4BV water with 4BV/h flow velocity eluting remove impurity, then with 5BV80% ethanol with 4BV/h flow velocity eluting, eluent is evaporated to without alcohol taste, it is dissolved in water and is settled to the volume identical with first time constant volume, treating that D101 macroporous adsorbent resin is further purified；
(4) loading repurity: by step (3) gained just purification of flavone solution with 1BV/h flow velocity loading to the D101 resin column handled well, d=10cm, h=60cm, the extracting liquid volume of 1BV=1.5 times, with 5BV40% ethanol with 4BV/h flow velocity eluting, starts to collect eluent from 2BV volume, after being evaporated to a small amount of volume, lyophilization, obtains flavone purified components, determination of color flavones content.
70%～85% described saponin is prepared by what method prepared:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder in the ratio powder of 10:1, adds 60 DEG C of petroleum ether of 8 times amount, ultrasonic degreasing 40min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) extract: weigh defat medical material, add 5 times amount 60% ethanol, soak 0.5h, circumfluence distillation 3 times, each 1h, extracting solution is evaporated to without alcohol taste, with distilled water, concentrated solution is settled to 0.2g crude drug/mL, after extraction, remaining medicinal residues retain, dry, polysaccharide to be extracted；
(3) saponin purification: by step (2) gained 0.2g crude drug/ml extracting solution with 1BV/h flow velocity loading to the DM130 resin column handled well, d=10cm, the extracting liquid volume of h=60cm, 1BV=1.5 times, after absorption 2h, with 3BV water with 4BV/h flow velocity eluting remove impurity, remove impurity is continued with 4BV/h flow velocity with 5BV20% ethanol, then with 5BV60% ethanol with 4BV/h flow velocity eluting saponin, the concentrated lyophilization of eluent, obtain saponin purification fractions, determination of color saponin content；
Or:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder in the ratio powder of 10:3, adds 75 DEG C of petroleum ether of 10 times amount, ultrasonic degreasing 50min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) extract: weigh defat medical material, add 12 times amount 70% ethanol, soak 0.8h, circumfluence distillation 3 times, each 1.5h, extracting solution is evaporated to without alcohol taste, with distilled water, concentrated solution is settled to 0.2g crude drug/mL, after extraction, remaining medicinal residues retain, dry, polysaccharide to be extracted；
(3) saponin purification: by step (2) gained 0.2g crude drug/ml extracting solution with 1BV/h flow velocity loading to the DM130 resin column handled well, d=10cm, the extracting liquid volume of h=60cm, 1BV=1.5 times, after absorption 2h, with 3BV water with 4BV/h flow velocity eluting remove impurity, remove impurity is continued with 4BV/h flow velocity with 5BV30% ethanol, then with 5BV70% ethanol with 4BV/h flow velocity eluting saponin, the concentrated lyophilization of eluent, obtain saponin purification fractions, determination of color saponin content；
Or:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder in the ratio powder of 10:5, adds 90 DEG C of petroleum ether of 12 times amount, ultrasonic degreasing 60min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) extract: weigh defat medical material, add 20 times amount 80% ethanol, soak 1h, circumfluence distillation 3 times, each 2h, extracting solution is evaporated to without alcohol taste, with distilled water, concentrated solution is settled to 0.2g crude drug/mL, after extraction, remaining medicinal residues retain, dry, polysaccharide to be extracted；
(3) saponin purification: by step (2) gained 0.2g crude drug/ml extracting solution with 1BV/h flow velocity loading to the DM130 resin column handled well, d=10cm, the extracting liquid volume of h=60cm, 1BV=1.5 times, after absorption 2h, with 3BV water with 4BV/h flow velocity eluting remove impurity, remove impurity is continued with 4BV/h flow velocity with 5BV40% ethanol, then with 5BV80% ethanol with 4BV/h flow velocity eluting saponin, the concentrated lyophilization of eluent, obtain saponin purification fractions, determination of color saponin content.
The effect of the present invention is further illustrated below by pharmacodynamic experiment.
1. materials and methods
1.1 materials and reagent
Experiment reagent is analytical pure, and water is ultra-pure water.STZ is purchased from Sigma Co., USA；Citric acid, sodium citrate are purchased from Yi Sheng bio tech ltd, Shanghai；Metformin hydrochloride is purchased from Shenzhen Haiwang Pharmaceutical Co., Ltd；Rat blood urea nitrogen quantification kit, Triglyceride Reagent box, T-CHOL test kit, low density lipoprotein, LDL test kit are purchased from Nanjing and build up Bioengineering Research Institute；Blood sugar test paper and blood glucose meter are Yicheng Biological Electronic Technology Co., Ltd., Beijing's product.
1.2 laboratory animals
SPF level health male SD rat is purchased from Jilin University's Experimental Animal Center, body weight (210 ± 20) g.
1.3 instrument and equipments
Centrifuge5810R high speed centrifuge (Eppendorf, the U.S.)；The multi-functional microplate reader of GENiosPro (Tecan, Switzerland)；Milli-QGradientA10 ultrapure water system (Millipore, the U.S.).
1.4 methods
1.4.1STZ type i diabetes rat model is set up in induction
Healthy SD male rat is randomly divided into two groups, blank group and modeling group, adaptability feeds 7d, water 16h is can't help in fasting, dosage is the STZ lumbar injection modeling of 65mg/kg, and STZ need to be dissolved in citrate buffer (pH=4.2) ice-cold for 0.1M before use, is made into the solution of 1% concentration, keep ice-cold state, and in 0.5h, complete injection.After one week, the fasting glucose >=16.7mM of rat is modeling success, random packet, makes blood glucose difference≤1mM between each group.
1.4.5 experiment packet and administration
Experiment packet and administration are detailed such as table 1, each group gastric infusion every day 1 time, successive administration 4 weeks.The front 37 DEG C of water-bath 20min of gavage.After the administration of each group terminates, remain medicinal liquid 4 DEG C storage, ready to be reused.
Table 1 experiment packet and administrations
Note: 10mL/kg, the i.e. every kg body weight gavage 10mL water of rat, positive drug or compatibility component solution；0.018g/kg, the i.e. every kg body weight gavage 0.018g positive drug of rat, 0.018g/kg is 10mL/kg；0.45g/kg, i.e. rat every kg body weight gavage 0.45g compatibility component, 0.45g/kg is 10mL/kg.
1.4.6 Indexs measure
(1) fasting glucose (FBG) measures: after gastric infusion, Rat Fast be can't help water 12h by 7d, 14d, 21d, 28d, and when the morning 8, the blood sampling of tail point measures FBG.
(2) renal tissue morphologic observation: rat place after death, dissects rapidly and takes out right side kidney (one section of osculum is cut off in side), and fixing in 10% formalin, HE dyes, tissues observed form under light microscopic.
(3) urine, Evaluation of blood test: after last administration terminates, water is can't help in fasting, collects the urine of 16h, records urine volume, and centrifugal under 4 DEG C of conditions collects supernatant；After metabolism of rat terminates, put to death, access blood, centrifugal collection serum after standing 2h under 4 DEG C of conditions.Take appropriate urine, measure urea nitrogen content in urine；Take appropriate serum, measure the content of triglyceride, T-CHOL, low density lipoprotein, LDL in rat blood serum.
1.4.7 statistical procedures
Experimental data withRepresenting, adopting SPSS statistical software to compare as between one factor analysis of variance group, significance level is with 0.05 and 0.01 for standard.
2. result
Diabetes rat FBG is affected by 2.1 compatibilities
The fasting glucose data of each experimental group 28d of table 2
Note: * and MC group compares, there were significant differences (P < 0.05)；* * and MC group compares, and has pole significant difference (P < 0.01), lower same.
Each administration group FBG gradually reduces along with continuing medication.After being administered 28 days, each administration group is obvious to the effect lowering FBG, and positive drug group has significance (P < 0.05), and 3 compatibility groups are respectively provided with pole significant difference (P < 0.01), wherein the hypoglycemic effect of 3:6:1 group is best, and under this group blood glucose tone pitch more than positive drug group.
The impact on kidneys of diabetic rats of 2.2 compatibilities
Kidney coloration result as shown in Figure 1, expand, renal tubules swelling, blood capillary obliteration, and mesangial cell increases, proximal tubular cell swelling and degeneration by model group glomerule, and cavity is formed.Each compatibility administration group and positive drug group, there are differences in glomerule degrees of expansion compared with model group, and blood capillary obliteration improves, and cavity reduces.Wherein 3:6:1 compatibility group is suppressing glomerule expansion, improves hair and blood pipe obliteration, reduces proximal convoluted tubule swelling aspect curative effect best, is better than other two compatibility groups or even metformin hydrochloride groups.
The impact on diabetes rat urine blood urea nitrogen of 2.3 compatibilities
Table 3 urine determination of urea nitrogen data
Compared with blank group, in model group rats urine, urea nitrogen content is significantly raised, and positive drug group, each compatibility administration group urea nitrogen content compared with model group substantially reduce, and all have statistical significance (P < 0.01), wherein 3:6:1 group therapeutic effect is optimum, and is better than positive drug group.
The impact on diabetes rat serum triglycerides, T-CHOL of 2.4 compatibilities
Each group rat blood serum triglyceride, T-CHOL measurement result are shown in accompanying drawing 2,3, compare with blank group, and model group rats serum triglycerides, T-CHOL are significantly raised.After positive drug group and the treatment of each compatibility administration group, rat blood serum triglyceride, total cholesterol level substantially reduce, compare with model group and there is pole significant difference (P < 0.01), and 3:6:1 compatibility group effect is better than other two compatibility groups, and similar with positive drug group therapeutic effect.
The impact on diabetes rat serum low-density LP of 2.5 compatibilities
Each group rat blood serum low-density LP determination result is shown in accompanying drawing 4, compares with blank group, and model group rats serum low-density LP is significantly raised.After positive drug group is treated with each compatibility administration group, rat blood serum low density lipoprotein, LDL content substantially reduces, and compares with model group and has pole significant difference (P < 0.01).Wherein 3:6:1 group therapeutic effect is optimum, and is better than positive drug group.
Learn that model group rats serum triglycerides, T-CHOL, low density lipoprotein, LDL are all significantly raised by the above results, illustrate that rat has occurred metabolism disorder of blood lipid, after being administered 28 days, each index is all improved, and illustrates that this invention has auxiliary lipid-lowering function.And rat blood sugar level substantially reduces after testing, wherein maximum with the change of 3:6:1 compatibility group, therefore can determine that this invention has the effect reducing blood-fat and blood sugar.
By above-described embodiment, the purpose of the present invention is reached by fully effective.It is familiar with the personage of this skill and should be understood that the present invention includes but not limited to accompanying drawing and content described in detailed description of the invention above.The function of any present invention of not necessarily departing from and the amendment of structural principle are intended to be included in the scope of claims.
Claims (4)
1. the pharmaceutical composition treating diabetes, it is characterised in that be prepared by what the medicine of parts by weight was made:
60%～70% polysaccharide 30～70 parts；
70%～85% saponin 25～60 parts；
55%～70% flavone 5～10 parts.
2. a kind of pharmaceutical composition treating diabetes according to claim 1, it is characterised in that: 60%～70% described polysaccharide is prepared by what method prepared:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder by the weight ratio powder of 10:1～5, adds 8～12 times amount 60-90 DEG C of petroleum ether, ultrasonic degreasing 40～60min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) Polyose extraction: weigh defat medical material, add 5～20 times amount 60%～80% ethanol, soak 0.5～1h, circumfluence distillation 3 times, each 1～2h, extracting solution is evaporated to without alcohol taste, and after extraction, remaining medicinal residues retain, dry, add 5～15 times amount distilled water, extract 60～120min, 10000r/min centrifugal 10min when 10 DEG C of magnetic agitation, collect supernatant；
(3) polysaccharide purification: 60%～80% ethanol precipitate with ethanol, stands overnight, and 4000r/min is centrifuged 10min, supernatant discarded, and precipitation distilled water fully redissolves, lyophilization, obtains polysaccharide purification component, determination of color polyoses content.
3. a kind of pharmaceutical composition treating diabetes according to claim 1, it is characterised in that: 55%～70% described flavone is prepared by what method prepared:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder by the weight ratio powder of 10:1～5, adds 60 DEG C～90 DEG C petroleum ether of 8～12 times amount, ultrasonic degreasing 40～60min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) extract: weigh defat medical material, add 5～20 times amount 60%～80% ethanol, soak 0.5～1h, circumfluence distillation 3 times, each 1～2h, extracting solution is evaporated to without alcohol taste, with distilled water, concentrated solution is settled to 0.2g crude drug/mL, after extraction, remaining medicinal residues retain, dry, polysaccharide to be extracted；
(3) loading just purification: by step (2) gained 0.2g crude drug/ml extracting solution with 1BV/h flow velocity loading to the polyamide resin column handled well, d=10cm, h=60cm, the extracting liquid volume of 1BV=1.5 times, with 4BV water with 4BV/h flow velocity eluting remove impurity, then with 5BV60%～80% ethanol with 4BV/h flow velocity eluting, eluent is evaporated to without alcohol taste, it is dissolved in water and is settled to the volume identical with first time constant volume, treating that D101 macroporous adsorbent resin is further purified；
(4) loading repurity: by step (3) gained just purification of flavone solution with 1BV/h flow velocity loading to the D101 resin column handled well, d=10cm, h=60cm, the extracting liquid volume of 1BV=1.5 times, with 5BV20%～40% ethanol with 4BV/h flow velocity eluting, starts to collect eluent from 2BV volume, after being evaporated to a small amount of volume, lyophilization, obtains flavone purified components, determination of color flavones content.
4. a kind of pharmaceutical composition treating diabetes according to claim 1, it is characterised in that: 70%～85% described saponin is prepared by what method prepared:
(1) medical material defat: Semen Trigonellae, the Radix Astragali become coarse powder by the weight ratio powder of 10:1～5, adds 8～12 times amount 60-90 DEG C of petroleum ether, ultrasonic degreasing 40～60min under 400w power, filters, and medicinal material coarse powder volatilizes petroleum ether, standby；
(2) extract: weigh defat medical material, add 5～20 times amount 60%～80% ethanol, soak 0.5～1h, circumfluence distillation 3 times, each 1～2h, extracting solution is evaporated to without alcohol taste, with distilled water, concentrated solution is settled to 0.2g crude drug/mL, after extraction, remaining medicinal residues retain, dry, polysaccharide to be extracted；
(3) saponin purification: by step (2) gained 0.2g crude drug/ml extracting solution with 1BV/h flow velocity loading to the DM130 resin column handled well, d=10cm, h=60cm, the extracting liquid volume of 1BV=1.5 times, after absorption 2h, with 3BV water with 4BV/h flow velocity eluting remove impurity, remove impurity is continued with 4BV/h flow velocity with 5BV20%～40% ethanol, again with 5BV60%～80% ethanol with 4BV/h flow velocity eluting saponin, the concentrated lyophilization of eluent, obtain saponin purification fractions, determination of color saponin content.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116622003A (en) * | 2023-07-03 | 2023-08-22 | 北京中医药大学 | Discovery preparation of traditional Chinese medicine material basis and application of traditional Chinese medicine material basis in reducing blood glucose and blood lipid |
| CN117903335A (en) * | 2024-01-25 | 2024-04-19 | 临沂大学 | Traditional Chinese medicine polysaccharide extract and application thereof in diabetes medical food |
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| CN1605357A (en) * | 2004-07-30 | 2005-04-13 | 北京星昊现代医药有限公司 | Fenugreek seed extract and its preparing process and application |
| CN1634463A (en) * | 2004-11-02 | 2005-07-06 | 华启洪 | Medicine for treating diabetes and production method thereof |
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| CN1605357A (en) * | 2004-07-30 | 2005-04-13 | 北京星昊现代医药有限公司 | Fenugreek seed extract and its preparing process and application |
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| CN116622003A (en) * | 2023-07-03 | 2023-08-22 | 北京中医药大学 | Discovery preparation of traditional Chinese medicine material basis and application of traditional Chinese medicine material basis in reducing blood glucose and blood lipid |
| CN116622003B (en) * | 2023-07-03 | 2024-04-05 | 北京中医药大学 | Discovery preparation of traditional Chinese medicine material basis and application of traditional Chinese medicine material basis in reducing blood glucose and blood lipid |
| CN117903335A (en) * | 2024-01-25 | 2024-04-19 | 临沂大学 | Traditional Chinese medicine polysaccharide extract and application thereof in diabetes medical food |
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Application publication date: 20160706 |