CN110507708B - Safflower Xiaoyao preparation for treating hyperplasia of mammary glands and preparation method and application thereof - Google Patents

Safflower Xiaoyao preparation for treating hyperplasia of mammary glands and preparation method and application thereof Download PDF

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CN110507708B
CN110507708B CN201910896093.3A CN201910896093A CN110507708B CN 110507708 B CN110507708 B CN 110507708B CN 201910896093 A CN201910896093 A CN 201910896093A CN 110507708 B CN110507708 B CN 110507708B
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parts
preparation
safflower
ethanol
solution
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CN110507708A (en
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陈梁
李小锋
刘厚权
夏淑英
胡吉忠
张俊英
张金良
周朝忠
毛金娣
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JIANGXI PUZHENG PHARMACEUTICAL CO Ltd
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JIANGXI PUZHENG PHARMACEUTICAL CO Ltd
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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
    • A61K36/06—Fungi, e.g. yeasts
    • A61K36/07—Basidiomycota, e.g. Cryptococcus
    • A61K36/076—Poria
    • 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)
    • A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
    • A61K36/232—Angelica
    • 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)
    • A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
    • A61K36/233—Bupleurum
    • 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)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • A61K36/284—Atractylodes
    • 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)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
    • A61K36/286—Carthamus (distaff thistle)
    • 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
    • A61K36/483—Gleditsia (locust)
    • 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
    • A61K36/484—Glycyrrhiza (licorice)
    • 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)
    • A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • A61K36/534—Mentha (mint)
    • 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)
    • A61K36/65—Paeoniaceae (Peony family), e.g. Chinese peony
    • 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)
    • A61K36/71—Ranunculaceae (Buttercup family), e.g. larkspur, hepatica, hydrastis, columbine or goldenseal
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/14—Drugs for genital or sexual disorders; Contraceptives for lactation disorders, e.g. galactorrhoea
    • 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, 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
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • 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
    • 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
    • A61K2236/55—Liquid-liquid separation; Phase separation

Abstract

The invention belongs to the preparation and application field of safflower Xiaoyao tablets, in particular to a safflower Xiaoyao preparation for treating hyperplasia of mammary glands and a preparation method thereof; the composite material comprises the following raw materials in parts by weight: 18-67 parts of angelica, 100-150 parts of white paeony root, 30-55 parts of bighead atractylodes rhizome, 135 parts of poria cocos, 110-160 parts of safflower, 20-35 parts of spina gleditsiae, 80-140 parts of bupleurum chinense, 45-85 parts of mint and 20-75 parts of liquorice; the safflower Xiaoyao preparation prepared by the specific preparation method can well play the effective role of the medicine, and has good regulation effect on female endocrine.

Description

Safflower Xiaoyao preparation for treating hyperplasia of mammary glands and preparation method and application thereof
Technical Field
The invention belongs to the field of preparation and application of safflower Xiaoyao tablets, and particularly relates to a safflower Xiaoyao preparation for treating hyperplasia of mammary glands and a preparation method and application thereof.
Background
Mastoplasia is also called as breast dysplasia, and common breast diseases with breast lumps, breast pain and nipple discharge as main symptoms are degenerative and progressive lesions of the structure of mammary ducts and lobules [1 ]. Hyperplasia of mammary glands is not only in the category of tumors, but also in the area of inflammation, but also in the structural changes of the lobules and ducts of the mammary glands [2 ]. According to the pathology, the traditional Chinese medicine can be divided into simple mammary epithelial hyperplasia, cystic hyperplasia of breast and mastopathy. The main clinical features of hyperplasia of mammary glands are breast lumps and periodic swelling pain of the breast. The mastosis accounts for more than 70 percent of all mastosis, is the most common mastosis of women of childbearing age, accounts for about 50 percent of the mastosis of women of childbearing age, has a trend of increasing prevalence year by year in recent years, and greatly influences the normal life of patients. The disease has the risk of developing into breast cancer to a certain extent, seriously threatens the health of women, and has important practical significance in researching the current situation of the drug therapy of the disease and effectively preventing and treating the disease.
Safflower is the dry cylindrical corolla of the plant safflower of the Compositae, also called safflower grass, safflower vegetable, Sichuan safflower and spiny safflower, because its leaves are similar to indigo blue and also called bluish flower, the name of bluish flower is originally seen in Ben Cao Tu Jing, which is called "bluish flower, i.e. safflower also". It is pungent and warm in nature. It has effects of promoting blood circulation, dredging channels, removing blood stasis, and relieving pain, and can be used for treating amenorrhea, dysmenorrhea, lochiorrhea, abdominal mass, thoracic obstruction, cardialgia, blood stasis, abdominal pain, pain in chest and hypochondrium, traumatic injury, and pyocutaneous disease with swelling and pain. Modern pharmacological research shows that safflower can slightly excite heart, reduce coronary resistance and increase coronary flow; protecting and improving myocardial ischemia, resisting arrhythmia, lowering blood pressure, and inhibiting platelet aggregation; improving the hypoxia tolerance. In addition, safflower also has the functions of sedation, analgesia, anti-inflammation and immunosuppression, and the decoction of the safflower has the excitation function on the smooth muscle of the uterus and the intestinal tract. The clinical application is mainly used for treating cardiovascular system diseases, gynecological diseases, nervous system diseases and other diseases, and the main effective components are safflower yellow, hydroxy safflower yellow, carthamin, safflower oil and the like. In the prior art, the traditional Chinese medicine composition of safflower is often used for preparing the medicine for treating hyperplasia of mammary glands.
Chinese patent application CN 104958693A discloses a traditional Chinese medicine for treating hyperplasia of mammary glands, which comprises the following raw material medicines: 5-15 parts of selfheal, 15-25 parts of safflower, 15-25 parts of salvia miltiorrhiza, 5-15 parts of spina gleditsiae, 10-20 parts of liquorice, 5-15 parts of angelica, 5-15 parts of red paeony root, 5-15 parts of radix bupleuri and 10-20 parts of radix curcumae.
Chinese patent application CN 1055623A discloses a Chinese patent medicine for treating female hyperplasia of mammary glands, which mainly comprises radix Paeoniae Rubra, rhizoma Ligustici Chuanxiong, Achyranthis radix, Saviae Miltiorrhizae radix, Carthami flos, semen Persicae, Sichuan hazelnut, pericarpium Citri Reticulatae viride, bupleuri radix, Prunellae Spica, Glycyrrhrizae radix, and Poria; the components for promoting blood circulation to remove blood stasis and soothing liver and relieving depression have the problem of eliminating lumps, but the application of the patent also has poor effect pertinence due to the limitation of the preparation method, and cannot well play the effective function of the medicine.
Research shows that after the safflower is taken for a long time, the safflower can cause serious damage to nerves, urinary system and digestive system, and is accompanied with adverse reactions such as skin allergy, anaphylactic shock and the like. For example, headache, rash, whelk, constipation, excessive internal heat, irregular menstruation, weakness all over the body, lower abdominal pain, gastrointestinal bleeding, unconsciousness, weak pulse and the like. These are a series of adverse reactions of taking safflower for a long time. Therefore, the reasonable preparation and application of safflower have important functions.
Disclosure of Invention
In order to overcome the technical problems, the invention provides the safflower Xiaoyao preparation for treating the hyperplasia of mammary glands and the preparation method thereof.
In order to achieve the above purpose, the technical scheme provided by the invention is as follows:
a safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following raw materials in parts by weight: 18-67 parts of angelica, 100-150 parts of white paeony root, 30-55 parts of bighead atractylodes rhizome, 135 parts of poria cocos, 110-160 parts of safflower, 20-35 parts of spina gleditsiae, 80-140 parts of bupleurum chinense, 45-85 parts of mint and 20-75 parts of liquorice;
preferably, the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following raw materials in parts by weight: 20-53 parts of angelica, 145 parts of white paeony root 125-;
preferably, the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following raw materials in parts by weight: 32 parts of angelica, 130 parts of white peony root, 50 parts of bighead atractylodes rhizome, 127 parts of poria cocos, 140 parts of safflower, 28 parts of spina gleditsiae, 110 parts of bupleurum chinense, 65 parts of mint and 55 parts of liquorice.
The invention also aims to provide a preparation method of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands, which comprises the following steps:
(1) pulverizing Carthami flos, Glycyrrhrizae radix, and bupleuri radix, adding water, mixing, and extracting to obtain water extract A and residue 1; adsorbing the water extract A by a macroporous adsorption resin chromatographic column, concentrating the residual water extract after adsorption to obtain a water extract B, and eluting the macroporous adsorption resin chromatographic column by an ethanol water solution to obtain an alcohol solution 1;
(2) extracting the residue 1 with ethanol to obtain ethanol extract, mixing the ethanol extract and the water extract B, and separating by silica gel column chromatography; eluting the silica gel chromatographic column with ethyl acetate, then eluting with dimethyl carbonate-ethanol mixed solution, concentrating and recrystallizing to obtain crystal C;
(3) extracting radix Angelicae sinensis, Atractylodis rhizoma, and herba Menthae with ethanol to obtain alcoholic solution 2 and residue 2;
(4) extracting the herb residue 2, the poria cocos, the spina gleditsiae and the white paeony root with water, and adding a flocculating agent for treatment to obtain an aqueous solution 1;
(5) mixing the alcoholic solution 1, the alcoholic solution 2 and the aqueous solution 1, adding the crystal C for dissolving, and concentrating to obtain a concentrated solution;
(6) and preparing the concentrated solution into a preparation, namely the safflower Xiaoyao preparation.
Preferably, in the step (1), the specific process of adding water and mixing for extraction is as follows: heating to 75-90 deg.C, extracting for 1-2h, boiling, and decocting for 20-30 min;
preferably, in step (1), the macroporous adsorption resin chromatographic column is: the upper layer is a chromatographic column consisting of AB-8, the middle layer is NKA and the lower layer is NKA-9, wherein the thickness ratio of the AB-8 to the NKA-9 is 1:1-2: 1-2;
preferably, in the step (1), the ethanol mass concentration of the ethanol aqueous solution is 30-50%;
preferably, in the step (2), the volume ratio of dimethyl carbonate to ethanol in the dimethyl carbonate-ethanol mixed solution is 1: 1-3;
preferably, in the step (3), the ethanol extraction is performed at a temperature of 40-60 ℃;
preferably, in step (4), the flocculant is chitosan or gelatin;
preferably, in the step (4), the flocculant is chitosan, and the pH value of the water extract is adjusted to 3-5 before the flocculation treatment, and then the chitosan is added;
preferably, in the step (4), the flocculant is gelatin, and the temperature of the water extract is increased to 50-60 ℃ during flocculation treatment, and then the gelatin is added;
preferably, in step (6), the preparation is a pharmaceutically conventional agent, including tablets, capsules, pills, and oral liquid.
Preferably, the step (1) further comprises a pretreatment of a macroporous adsorption resin chromatographic column, and the specific process comprises soaking the chromatographic column in ethanol for 20-30min, and then soaking in water for 2-3 h.
The invention also aims to provide application of the safflower Xiaoyao preparation or the safflower Xiaoyao preparation prepared by the preparation method of the safflower Xiaoyao preparation in preparing a medicine for treating hyperplasia of mammary glands.
Compared with the prior art, the invention has the technical advantages that:
(1) the safflower Xiaoyao preparation has better treatment effect on treating hyperplasia of mammary glands by improving the dosage of each raw material in the safflower Xiaoyao preparation;
(2) in the preparation process of the safflower Xiaoyao preparation, the safflower, the bupleurum root of bamboo leaf and the liquorice are mixed and extracted to play the role of promoting the dissolution of effective components, thereby being more beneficial to playing the synergistic effect of the effective components; the extract is separated by macroporous adsorption resin and then is separated by silica gel column chromatography, so that impurities in the medicament can be well removed, the effective components are purified, and meanwhile, the side effects of the medicament are reduced;
(3) the angelica, the white atractylodes rhizome and the mint are subjected to ethanol extraction, so that volatile oil in the angelica, the white atractylodes rhizome and the mint can be extracted, and the volatile oil and the safflower component can be used together to exert a synergistic effect, so that the effects of soothing the liver, regulating qi, promoting blood circulation, removing blood stasis, softening hardness, dissipating stagnation, regulating endocrine and the like are achieved;
(4) through the improvement of the preparation method, the safflower Xiaoyao preparation has better treatment effect on hyperplasia of mammary glands, simultaneously reduces the impurity content in components such as safflower and the like, and improves the efficacy of the effective components of the safflower Xiaoyao preparation.
Detailed Description
Example 1
In the embodiment, the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following raw materials in parts by weight: 32 parts of angelica, 130 parts of white peony root, 50 parts of bighead atractylodes rhizome, 127 parts of poria cocos, 140 parts of safflower, 28 parts of spina gleditsiae, 110 parts of bupleurum chinense, 65 parts of mint and 55 parts of liquorice.
The preparation method of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following steps:
(1) pulverizing Carthami flos, Glycyrrhrizae radix, and bupleuri radix, adding water, extracting at 80 deg.C for 1.5 hr, boiling, and decocting for 25 min; obtaining water extract A and medicine dregs 1;
taking a macroporous adsorption resin chromatographic column: the upper layer is a chromatographic column consisting of AB-8, the middle layer is NKA and the lower layer is NKA-9, wherein the thickness ratio of the AB-8 to the NKA-9 is 1:2: 1; soaking the chromatographic column in ethanol for 25min, and soaking in water for 2.5 h;
adsorbing the water extract A by using a macroporous adsorption resin chromatographic column, concentrating the residual water extract after adsorption to obtain a water extract B, and eluting the macroporous adsorption resin chromatographic column by using 40% ethanol water solution by mass concentration to obtain an alcohol solution 1;
(2) extracting the residue 1 with ethanol to obtain ethanol extract, mixing the ethanol extract and the water extract B, and separating by silica gel column chromatography; eluting the silica gel chromatographic column with ethyl acetate, then eluting with dimethyl carbonate-ethanol mixed solution with the volume ratio of 1:2, concentrating and recrystallizing to obtain crystal C;
(3) extracting radix Angelicae sinensis, Atractylodis rhizoma, and herba Menthae with ethanol at 50 deg.C to obtain alcoholic solution 2 and residue 2;
(4) extracting the residue 2, Poria, spina Gleditsiae and radix Paeoniae alba with water, adjusting pH to 3, and adding chitosan to obtain water solution 1;
(5) mixing the alcoholic solution 1, the alcoholic solution 2 and the aqueous solution 1, adding the crystal C for dissolving, and concentrating to obtain a concentrated solution;
(6) and (4) preparing the concentrated solution into tablets to obtain the safflower Xiaoyao tablets.
Example 2
A safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following raw materials in parts by weight: 18 parts of angelica, 100 parts of white peony root, 55 parts of bighead atractylodes rhizome, 135 parts of poria cocos, 110 parts of safflower, 20 parts of spina gleditsiae, 140 parts of bupleurum chinense, 85 parts of mint and 20 parts of liquorice;
the preparation method of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following steps:
(1) pulverizing Carthami flos, Glycyrrhrizae radix, and bupleuri radix, adding water, extracting at 75 deg.C for 1 hr, boiling, and decocting for 30 min; obtaining water extract A and medicine dregs 1;
taking a macroporous adsorption resin chromatographic column: the upper layer is a chromatographic column consisting of AB-8, the middle layer is NKA and the lower layer is NKA-9, wherein the thickness ratio of the AB-8 to the NKA-9 is 1:1: 1; soaking the chromatographic column in ethanol for 20min, and soaking in water for 2 h;
adsorbing the water extract A by using a macroporous adsorption resin chromatographic column, concentrating the residual water extract after adsorption to obtain a water extract B, and eluting the macroporous adsorption resin chromatographic column by using an ethanol water solution with the mass concentration of 30% to obtain an alcohol solution 1;
(2) extracting the residue 1 with ethanol to obtain ethanol extract, mixing the ethanol extract and the water extract B, and separating by silica gel column chromatography; eluting the silica gel chromatographic column with ethyl acetate, then eluting with dimethyl carbonate-ethanol mixed solution with the volume ratio of 1:1, concentrating and recrystallizing to obtain crystal C;
(3) extracting radix Angelicae sinensis, Atractylodis rhizoma, and herba Menthae with ethanol at 40 deg.C to obtain alcoholic solution 2 and residue 2;
(4) extracting the dregs 2, the tuckahoe, the spina gleditsiae and the white paeony root with water, raising the temperature of the water extract to 50 ℃, and then adding gelatin for treatment to obtain an aqueous solution 1;
(5) mixing the alcoholic solution 1, the alcoholic solution 2 and the aqueous solution 1, adding the crystal C for dissolving, and concentrating to obtain a concentrated solution;
(6) and (4) preparing the concentrated solution into capsules to obtain the safflower Xiaoyao capsules.
Example 3
A safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following raw materials in parts by weight: 67 parts of angelica, 150 parts of white paeony root, 30 parts of bighead atractylodes rhizome, 105 parts of tuckahoe, 160 parts of safflower, 35 parts of spina gleditsiae, 80 parts of bupleurum chinense, 45 parts of mint and 75 parts of liquorice;
the preparation method of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following steps:
(1) pulverizing Carthami flos, Glycyrrhrizae radix, and bupleuri radix, adding water, extracting at 90 deg.C for 2 hr, boiling, and decocting for 20 min; obtaining water extract A and medicine dregs 1;
taking a macroporous adsorption resin chromatographic column: the upper layer is a chromatographic column consisting of AB-8, the middle layer is NKA and the lower layer is NKA-9, wherein the thickness ratio of the AB-8 to the NKA-9 is 1:2: 2; soaking the chromatographic column in ethanol for 30min, and soaking in water for 3 h;
adsorbing the water extract A by using a macroporous adsorption resin chromatographic column, concentrating the residual water extract after adsorption to obtain a water extract B, and eluting the macroporous adsorption resin chromatographic column by using 50% ethanol water solution by mass concentration to obtain an alcohol solution 1;
(2) extracting the residue 1 with ethanol to obtain ethanol extract, mixing the ethanol extract and the water extract B, and separating by silica gel column chromatography; eluting the silica gel chromatographic column with ethyl acetate, then eluting with dimethyl carbonate-ethanol mixed solution with the volume ratio of 1:3, concentrating and recrystallizing to obtain crystal C;
(3) extracting radix Angelicae sinensis, Atractylodis rhizoma, and herba Menthae with ethanol at 40 deg.C to obtain alcoholic solution 2 and residue 2;
(4) extracting the residue 2, Poria, spina Gleditsiae and radix Paeoniae alba with water, adjusting pH to 5, and adding chitosan to obtain water solution 1;
(5) mixing the alcoholic solution 1, the alcoholic solution 2 and the aqueous solution 1, adding the crystal C for dissolving, and concentrating to obtain a concentrated solution;
(6) making the concentrated solution into pill to obtain the safflower ease pill.
Example 4
A safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following raw materials in parts by weight: 20 parts of angelica, 125 parts of white paeony root, 45 parts of bighead atractylodes rhizome, 130 parts of tuckahoe, 153 parts of safflower, 30 parts of spina gleditsiae, 95 parts of bupleurum chinense, 55 parts of mint and 35 parts of liquorice;
the preparation method of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following steps:
(1) pulverizing Carthami flos, Glycyrrhrizae radix, and bupleuri radix, adding water, extracting at 75-90 deg.C for 1-2 hr, boiling for 20-30 min; obtaining water extract A and medicine dregs 1;
taking a macroporous adsorption resin chromatographic column: the upper layer is a chromatographic column consisting of AB-8, the middle layer is NKA and the lower layer is NKA-9, wherein the thickness ratio of the AB-8 to the NKA-9 is 1:1: 2; soaking the chromatographic column in ethanol for 20-30min, and soaking in water for 2-3 h;
adsorbing the water extract A by using a macroporous adsorption resin chromatographic column, concentrating the residual water extract after adsorption to obtain a water extract B, and eluting the macroporous adsorption resin chromatographic column by using 50% ethanol water solution by mass concentration to obtain an alcohol solution 1;
(2) extracting the residue 1 with ethanol to obtain ethanol extract, mixing the ethanol extract and the water extract B, and separating by silica gel column chromatography; eluting the silica gel chromatographic column with ethyl acetate, then eluting with dimethyl carbonate-ethanol mixed solution with the volume ratio of 1:3, concentrating and recrystallizing to obtain crystal C;
(3) extracting radix Angelicae sinensis, Atractylodis rhizoma, and herba Menthae with ethanol at 60 deg.C to obtain alcoholic solution 2 and residue 2;
(4) extracting the residue 2, Poria, spina Gleditsiae and radix Paeoniae alba with water, heating the water extractive solution to 60 deg.C, and adding gelatin to obtain water solution 1;
(5) mixing the alcoholic solution 1, the alcoholic solution 2 and the aqueous solution 1, adding the crystal C for dissolving, and concentrating to obtain a concentrated solution;
(6) making the concentrated solution into oral liquid to obtain Carthami flos XIAOYAO oral liquid.
Example 5
A safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following raw materials in parts by weight: 53 parts of angelica, 145 parts of white paeony root, 50 parts of bighead atractylodes rhizome, 125 parts of tuckahoe, 123 parts of safflower, 25 parts of spina gleditsiae, 120 parts of bupleurum chinense, 80 parts of mint and 65 parts of liquorice;
(1) pulverizing Carthami flos, Glycyrrhrizae radix, and bupleuri radix, adding water, extracting at 80 deg.C for 2 hr, boiling, and decocting for 30 min; obtaining water extract A and medicine dregs 1;
taking a macroporous adsorption resin chromatographic column: the upper layer is a chromatographic column consisting of AB-8, the middle layer is NKA and the lower layer is NKA-9, wherein the thickness ratio of the AB-8 to the NKA-9 is 1:2: 1; soaking the chromatographic column in ethanol for 30min, and soaking in water for 3 h;
adsorbing the water extract A by using a macroporous adsorption resin chromatographic column, concentrating the residual water extract after adsorption to obtain a water extract B, and eluting the macroporous adsorption resin chromatographic column by using 50% ethanol water solution by mass concentration to obtain an alcohol solution 1;
(2) extracting the residue 1 with ethanol to obtain ethanol extract, mixing the ethanol extract and the water extract B, and separating by silica gel column chromatography; eluting the silica gel chromatographic column with ethyl acetate, then eluting with dimethyl carbonate-ethanol mixed solution with the volume ratio of 1:2, concentrating and recrystallizing to obtain crystal C;
(3) extracting radix Angelicae sinensis, Atractylodis rhizoma, and herba Menthae with ethanol at 60 deg.C to obtain alcoholic solution 2 and residue 2;
(4) extracting the residue 2, Poria, spina Gleditsiae and radix Paeoniae alba with water, adjusting pH of the water extractive solution to 4, and adding chitosan to obtain water solution 1;
(5) mixing the alcoholic solution 1, the alcoholic solution 2 and the aqueous solution 1, adding the crystal C for dissolving, and concentrating to obtain a concentrated solution;
(6) and (4) preparing the concentrated solution into tablets to obtain the safflower Xiaoyao tablets.
Comparative example 1
The difference compared to example 1 is only the amount of safflower.
In the embodiment, the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following raw materials in parts by weight: 32 parts of angelica, 130 parts of white peony root, 50 parts of bighead atractylodes rhizome, 127 parts of poria cocos, 200 parts of safflower, 28 parts of spina gleditsiae, 110 parts of bupleurum chinense, 65 parts of mint and 55 parts of liquorice.
The preparation method of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands is the same as the example 1.
Comparative example 2
The difference compared to example 1 is only the amount of safflower.
In the embodiment, the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following raw materials in parts by weight: 32 parts of angelica, 130 parts of white peony root, 50 parts of bighead atractylodes rhizome, 127 parts of poria cocos, 80 parts of safflower, 28 parts of spina gleditsiae, 110 parts of bupleurum chinense, 65 parts of mint and 55 parts of liquorice.
The preparation method of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands is the same as the example 1.
Comparative example 3
The difference from the embodiment 1 is that the safflower, the bupleurum root and the liquorice are respectively and independently extracted;
in the embodiment, the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following raw materials in parts by weight: 32 parts of angelica, 130 parts of white peony root, 50 parts of bighead atractylodes rhizome, 127 parts of poria cocos, 140 parts of safflower, 28 parts of spina gleditsiae, 110 parts of bupleurum chinense, 65 parts of mint and 55 parts of liquorice.
The preparation method of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following steps:
(1) pulverizing Carthami flos, Glycyrrhrizae radix, and bupleuri radix respectively, adding water, extracting, heating to 80 deg.C for 1.5 hr, boiling, and decocting for 25 min; filtering to obtain Carthami flos extract, Glycyrrhrizae radix extract, bupleuri radix extract, Carthami flos residue, Glycyrrhrizae radix residue, and bupleuri radix residue; mixing the safflower extract, the liquorice extract and the bamboo leaf and bupleurum extract to obtain a water extract A;
taking a macroporous adsorption resin chromatographic column: the upper layer is a chromatographic column consisting of AB-8, the middle layer is NKA and the lower layer is NKA-9, wherein the thickness ratio of the AB-8 to the NKA-9 is 1:2: 1; soaking the chromatographic column in ethanol for 25min, and soaking in water for 2.5 h;
adsorbing the water extract A by using a macroporous adsorption resin chromatographic column, concentrating the residual water extract after adsorption to obtain a water extract B, and eluting the macroporous adsorption resin chromatographic column by using 40% ethanol water solution by mass concentration to obtain an alcohol solution 1;
(2) extracting Carthami flos residue, Glycyrrhrizae radix residue, and bupleuri radix residue with ethanol respectively, mixing the ethanol extracts of the three materials to obtain mixed ethanol extract, mixing the mixed ethanol extract with water extract B, and separating by silica gel column chromatography; eluting the silica gel chromatographic column with ethyl acetate, then eluting with dimethyl carbonate-ethanol mixed solution with the volume ratio of 1:2, concentrating and recrystallizing to obtain crystal C;
(3) - (6) same as in example 1.
Comparative example 4
Compared to example 1, no flocculant was added.
In this example, the raw materials of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands are the same as those in example 1.
The preparation method of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following steps:
(1) - (3) same as in example 1;
(4) extracting the residue 2, Poria, spina Gleditsiae and radix Paeoniae alba with water, and filtering to obtain water solution 1;
(5) - (6) same as in example 1.
Comparative example 5
The extraction method of the active ingredient was different from that of example 1.
In this example, the materials of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands are the same as those in example 1.
The preparation method of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following steps:
(1) pulverizing Carthami flos, Glycyrrhrizae radix, bupleuri radix, Poria, spina Gleditsiae and radix Paeoniae alba, adding water, extracting, heating to 80 deg.C for 1.5 hr, boiling, and decocting for 25 min; obtaining water extract A and medicine dregs 1;
(2) extracting the residue 1, radix Angelicae sinensis, Atractylodis rhizoma, and herba Menthae with ethanol to obtain ethanol extract;
(3) mixing the alcohol extract and the water extract A; concentrating to obtain concentrated solution;
(4) and (4) preparing the concentrated solution into tablets to obtain the safflower Xiaoyao tablets.
Examples of effects
(1) Rat mammary gland hyperplasia modeling
120 female rats with the weight of 200 +/-20 g are taken, and female rats are provided with SPF (specific pathogen free) grades and female animals from the breeding farm of Qinglongshan animals in Jiangning district of Nanjing city (SCXK (Su) 2017-.
Randomly divided into 12 groups of 10 animals each, examples 1-5 and comparative examples 1-5, model group and blank control group, respectively, and each rat in the blank control group was injected with 0.9% sodium chloride injection 0.2ml per day into the posterior thigh muscle for 30 days continuously, and the remaining 11 groups were injected with 0.5 mg/kg of estradiol benzoate injection diluted with olive oil (1: 9)-15 mg/kg progesterone injection diluted with olive oil (1:19) for 25 days continuously and on 26 days-1And 5d in succession.
(2) Determination of the level of E2, P
The safflower Xiaoyao preparations prepared in examples 1 to 5 and comparative examples 1 to 5 with the effective content of 0.25g/kg are respectively fed on the 5 th day of the molding, and the same amount of normal saline is fed to the blank control group. Continuously taking the medicine for 30 days, 35 days, 1h after the last administration, taking blood from orbit (centrifuging for 5min at 4000 rpm after 30min, taking serum), and measuring the levels of E2 and P in the serum by a radioimmunoassay. The results are shown in Table 1:
TABLE 1 serum E of rats2And variation of P content
Figure 518257DEST_PATH_IMAGE001
Remarking: there were significant differences (P < 0.05) between the different letters in the same column.
Compared with the control group, the influence on the levels of the E2 and the P in the serum of the rat is shown that the level of the E2 in the serum of the rat in the model group is obviously increased, and the level of the P in the serum is obviously reduced, which indicates that the model building is successful.
Compared with a model group, the safflower Xiaoyao preparation of the embodiment 1 to 5 of the invention can obviously reduce the level of E2 in the serum of rats and obviously increase the level of P in the serum, thereby showing that the safflower Xiaoyao preparation of the invention has better treatment effect on hyperplasia of mammary glands, and the test data of the comparative examples 1 to 2 show that: the dosage of the raw materials of the safflower Xiaoyao preparation has important influence on the effect; meanwhile, from the test data of comparative example-5, it can be seen that: the extraction and separation method of the effective components also has influence on the effect.
(3) Morphological changes of mammary gland tissue
After the serum is measured, all groups of rats are killed by a cervical vertebra removal method, the complete breasts are taken down, fixed by 10 percent formaldehyde solution, subjected to conventional flaking, HE staining, observed under a light microscope and subjected to pathological analysis; the lobules of mammary gland and acinus and lobular interstitium (each pathological section takes 3 different visual fields) of the lobules of mammary gland and lobules of the rats in each group are measured by stereo metrology, and the observation and measurement indexes are the volume of the lobules of mammary gland and the density (Vv) of interstitium and the specific surface (S) of the acinus of mammary gland. The bulk density is the ratio (%) of the reaction observed body to the reference system, and the specific surface area is the ratio (%) of the limiting membrane to the volume of the reaction sphere or bundle-like structure. The results are shown in Table 2:
TABLE 2 morphological changes in mammary lobules of rats
Figure 140737DEST_PATH_IMAGE002
Remarking: there were significant differences (P < 0.05) between the different letters in the same column.
The experimental data show that compared with the control group, the density of the mammary lobular body and the acinus specific surface of the mammary gland of the model group are obviously increased; the plasmid density between the leaflets is obviously reduced, which indicates that the molding is successful.
Compared with a model group, the safflower Xiaoyao preparation of the embodiment 1 to 5 of the invention can obviously reduce the density of the lobular body of the mammary gland and the specific surface of the acinus, and obviously increase the density of the interstitial body of the lobule; therefore, the safflower Xiaoyao preparation has better treatment effect on hyperplasia of mammary glands, and the test data of the comparative examples 1-2 show that: the dosage of the raw materials of the safflower Xiaoyao preparation has important influence on the effect; meanwhile, from the test data of comparative example-5, it can be seen that: the extraction and separation method of the effective components also has influence on the effect.
The above detailed description is specific to one possible embodiment of the present invention, and the embodiment is not intended to limit the scope of the present invention, and all equivalent implementations or modifications without departing from the scope of the present invention should be included in the technical scope of the present invention.

Claims (9)

1. The safflower Xiaoyao preparation for treating hyperplasia of mammary glands is characterized by comprising the following raw materials in parts by weight: 18-67 parts of angelica, 100-150 parts of white paeony root, 30-55 parts of bighead atractylodes rhizome, 135 parts of poria cocos, 110-160 parts of safflower, 20-35 parts of spina gleditsiae, 80-140 parts of bupleurum chinense, 45-85 parts of mint and 20-75 parts of liquorice; the preparation method of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands comprises the following steps:
(1) pulverizing Carthami flos, Glycyrrhrizae radix, and bupleuri radix, adding water, mixing, and extracting to obtain water extract A and residue 1; adsorbing the water extract by macroporous adsorption resin chromatographic column, concentrating the residual water extract after adsorption to obtain water extract B, and eluting the macroporous adsorption resin chromatographic column with ethanol water solution to obtain alcohol solution 1;
(2) extracting the residue 1 with ethanol to obtain ethanol extract, mixing the ethanol extract and the water extract B, and separating by silica gel column chromatography; eluting the silica gel chromatographic column with ethyl acetate, then eluting with dimethyl carbonate-ethanol mixed solution, concentrating and recrystallizing to obtain crystal C;
(3) extracting radix Angelicae sinensis, Atractylodis rhizoma, and herba Menthae with ethanol to obtain alcoholic solution 2 and residue 2;
(4) extracting the herb residue 2, the poria cocos, the spina gleditsiae and the white paeony root with water, and adding a flocculating agent for treatment to obtain an aqueous solution 1;
(5) mixing the alcoholic solution 1, the alcoholic solution 2 and the aqueous solution 1, adding the crystal C for dissolving, and concentrating to obtain a concentrated solution;
(6) preparing the concentrated solution into a preparation to obtain a safflower Xiaoyao preparation;
the macroporous adsorption resin chromatographic column comprises: the upper layer is a chromatographic column consisting of AB-8, the middle layer is NKA and the lower layer is NKA-9, wherein the thickness ratio of the AB-8 to the NKA-9 is 1:1-2: 1-2.
2. The safflower Xiaoyao preparation as claimed in claim 1, which comprises the following raw materials by weight: 20-53 parts of angelica, 145 parts of white paeony root 125-.
3. The safflower Xiaoyao preparation as claimed in claim 1, which comprises the following raw materials by weight: 32 parts of angelica, 130 parts of white peony root, 50 parts of bighead atractylodes rhizome, 127 parts of poria cocos, 140 parts of safflower, 28 parts of spina gleditsiae, 110 parts of bupleurum chinense, 65 parts of mint and 55 parts of liquorice.
4. The preparation method of the safflower Xiaoyao preparation for treating hyperplasia of mammary glands according to any one of claims 1 to 3, which comprises the following steps:
(1) pulverizing Carthami flos, Glycyrrhrizae radix, and bupleuri radix, adding water, mixing, and extracting to obtain water extract A and residue 1; adsorbing the water extract by macroporous adsorption resin chromatographic column, concentrating the residual water extract after adsorption to obtain water extract B, and eluting the macroporous adsorption resin chromatographic column with ethanol water solution to obtain alcohol solution 1;
(2) extracting the residue 1 with ethanol to obtain ethanol extract, mixing the ethanol extract and the water extract B, and separating by silica gel column chromatography; eluting the silica gel chromatographic column with ethyl acetate, then eluting with dimethyl carbonate-ethanol mixed solution, concentrating and recrystallizing to obtain crystal C;
(3) extracting radix Angelicae sinensis, Atractylodis rhizoma, and herba Menthae with ethanol to obtain alcoholic solution 2 and residue 2;
(4) extracting the herb residue 2, the poria cocos, the spina gleditsiae and the white paeony root with water, and adding a flocculating agent for treatment to obtain an aqueous solution 1;
(5) mixing the alcoholic solution 1, the alcoholic solution 2 and the aqueous solution 1, adding the crystal C for dissolving, and concentrating to obtain a concentrated solution;
(6) preparing the concentrated solution into a preparation to obtain a safflower Xiaoyao preparation;
the macroporous adsorption resin chromatographic column comprises: the upper layer is a chromatographic column consisting of AB-8, the middle layer is NKA and the lower layer is NKA-9, wherein the thickness ratio of the AB-8 to the NKA-9 is 1:1-2: 1-2.
5. The preparation method of the safflower Xiaoyao preparation for treating mammary gland hyperplasia according to claim 4, wherein in the step (1), the specific process of adding water for mixing and extracting is as follows: heating to 75-90 deg.C, extracting for 1-2 hr, boiling, and decocting for 20-30 min.
6. The method for preparing the safflower Xiaoyao preparation for treating mammary gland hyperplasia according to claim 4, wherein in the step (2), the volume ratio of dimethyl carbonate to ethanol in the dimethyl carbonate-ethanol mixed solution is 1: 1-3.
7. The method for preparing safflower Xiaoyao preparation for treating mammary gland hyperplasia according to claim 4, wherein the ethanol extraction is performed at 40-60 ℃ in the step (3).
8. The method for preparing the safflower Xiaoyao preparation for treating mammary gland hyperplasia according to claim 4, wherein in the step (4), the flocculating agent is chitosan or gelatin.
9. Use of the safflower Xiaoyao preparation for treating mammary gland hyperplasia according to any one of claims 1 to 3 or the preparation method of the safflower Xiaoyao preparation according to any one of claims 4 to 8 in the preparation of a medicament for treating mammary gland hyperplasia.
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