CN106267165A - HGH and IGF 1 liposome complex capsule and preparation method thereof - Google Patents
HGH and IGF 1 liposome complex capsule and preparation method thereof Download PDFInfo
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- CN106267165A CN106267165A CN201610765714.0A CN201610765714A CN106267165A CN 106267165 A CN106267165 A CN 106267165A CN 201610765714 A CN201610765714 A CN 201610765714A CN 106267165 A CN106267165 A CN 106267165A
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- 239000002775 capsule Substances 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title claims abstract description 26
- 101150088952 IGF1 gene Proteins 0.000 title abstract 4
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- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 21
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- 235000010469 Glycine max Nutrition 0.000 claims abstract description 19
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 88
- 108090000723 Insulin-Like Growth Factor I Proteins 0.000 claims description 80
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- 239000000706 filtrate Substances 0.000 claims description 60
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 48
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims description 38
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- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 24
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 21
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- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 20
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- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims description 14
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- 239000000787 lecithin Substances 0.000 claims description 14
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 10
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- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Substances N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 3
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- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
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- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
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- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-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
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/30—Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2
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- 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
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- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/288—Taraxacum (dandelion)
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- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/75—Rutaceae (Rue family)
- A61K36/752—Citrus, e.g. lime, orange or lemon
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K36/18—Magnoliophyta (angiosperms)
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- A61K36/82—Theaceae (Tea family), e.g. camellia
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- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8969—Polygonatum (Solomon's seal)
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- A61K38/011—Hydrolysed proteins; Derivatives thereof from plants
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/22—Hormones
- A61K38/27—Growth hormone [GH], i.e. somatotropin
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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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- A61K2236/331—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
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- 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
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
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- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Botany (AREA)
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- Alternative & Traditional Medicine (AREA)
- Medical Informatics (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Endocrinology (AREA)
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Abstract
The invention discloses a kind of HGH and IGF 1 liposome complex capsule and preparation method thereof, HGH, IGF 1 and soybean polypeptide are as primary raw material, with green tea, Chinese medicine extract combination, certain method is used to make liposome, make capsule preparations again, overcoming HGH and IGF 1 can only be by injection system just absorbent shortcoming, the capsule prepared is possible not only to the growth promotion that keeps healthy, and immunity, blood sugar lowering can be improved, having the effect for the treatment of poliomyelitis, drug effect and bioavailability are higher simultaneously.
Description
Technical field
The present invention relates to a kind of HGH and IGF-1 liposome complex capsule and preparation method thereof, belong to field of medicine preparations.
Background technology
HGH is human growth hormone, and human growth hormone is a kind of protein growth factor of pituitary secretion, is human body
Most important hormone compound, has and stimulates and the overall growth of regulation human body, and skeleton and muscle growth, vitals are sent out
Educate, the most important effect such as function of immune system.By stimulating various somatomedin.Growth hormone can promote the life of various kinds of cell
Length and differentiation, thus promote various tissue, the growth of organ, and suppress simultaneously or reverse human aging process.Human development
Peak period, i.e. ten how old by 20 years old, and in human body, auxin secretory volume peaks.
Growing up along with health enters, tumor growth element secretory volume declines year by year, live acoustic, within every ten years, declines percent
14, during by 80 years old, about 5 percent when tumor growth element secretory volume only has 20 years old.Along with tumor growth element is secreted
The reduction of amount, human body progresses into aging state, old and feeble various symptoms, as hoary, immunity and physically-draining occurs,
Libido declines, and fat increases, and cardiovascular regulating liver-QI pulmonary function reduces, and wrinkle, the bone loose easy generation fracture of energy etc. occurs in Dermatochalasia.
The full name of IGF-1 is called number growth factor of para-insulin.IGF-1 also referred to as " somatomedin ", is a kind of
On molecular structure with insulin type as polypeptide protein material.IGF-1 is persistently carried out in the growth of baby with in adult body
On anabolic action significant.
Genetic engineering recombinant protein is one of high-tech primary product of modern biological project.From first genetic engineering
Pharmaceutical grade protein in early eighties successfully as can be mass-produced commercialization medicine listing since, engineered protein
Matter has become as one of main product of modern medicine pharmaceutical market as bioengineered pharmaceuticals.For a long time, protein product
Being generally believed can only could realize effectively absorption by injection system by medical circle always, oral way is difficult to avoid that gastronintestinal system
Degraded, thus lose activity.
Summary of the invention
It is an object of the invention to provide a kind of HGH and IGF-1 liposome complex capsule and preparation method thereof, HGH, IGF-1
With soybean polypeptide as primary raw material, and the combination of green tea, Chinese medicine extract, use certain method to make liposome, then make
Capsule preparations, overcoming HGH and IGF-1 can only be by injection system just absorbent shortcoming, and prepared capsule is possible not only to
Health care growth promotion, and immunity, blood sugar lowering can be improved, there are the effect for the treatment of poliomyelitis, simultaneously drug effect and biology profit
Expenditure is higher.
HGH and IGF-1 liposome, is made up of the raw material of following masses number: HGH15-25 part, IGF-1 15-25 part, big
Bean polypeptide 14-18 part, green tea 5-10 part, Fructus Citri Sarcodactylis 5-10 part, Herba Taraxaci 2-4 part, Rhizoma Polygonati Odorati 2-4 part, lecithin 30-40 part, inositol
Phosphatidase 3 0-40 part, cholesterol 50-60 part, polyoxyethylene sorbitan monoleate 20-30 part, ethyl acetate 100-140 part, isopropanol 100-140
Part, phosphate buffer 1 80-220 part.
HGH and IGF-1 liposome, preparation method is:
(1) cholesterol of lecithin and half formula ratio is dissolved in ethyl acetate, mix homogeneously, removing is evaporated under reduced pressure molten
Agent, obtains immobilized artificial membrane I；
(2) cholesterol of lipositol He second half formula ratio is dissolved in isopropanol, mix homogeneously, is evaporated under reduced pressure and removes
Solvent, obtains immobilized artificial membrane II；
(3) immobilized artificial membrane I and immobilized artificial membrane II being added to the phosphate buffer of half formula ratio, stirring makes immobilized artificial membrane complete
Fully hydrated, make blank liposome suspension；
(4) HGH, IGF-1, soybean polypeptide and Chinese medicine extract are dissolved in the phosphate buffer of second half formula ratio,
Add blank liposome suspension, obtain HGH and IGF-1 liposome turbid liquor；
(5) HGH and IGF-1 liposome turbid liquor is spray-dried, obtains HGH and IGF-1 liposome.
Wherein, the preparation method of Chinese medicine extract is:
(1) green tea is added distilled water according to the ratio that solid-to-liquid ratio is 1g:16-20ml, soaks 2-3 hour at 80-90 DEG C,
Filter, obtain filtrate a；
(2) Herba Taraxaci is added to filtrate a, 200W supersound extraction 30-40 minute, filter, obtain filtrate b, standby；
(3) Fructus Citri Sarcodactylis is pulverized, adds 75% ethanol ultrasonic extraction 40-50 minute according to the ratio that solid-to-liquid ratio is 1g:9-11ml,
Filter, obtain filtrate c；
(4) filtrate b and filtrate c is merged, obtain mixed liquor, extract with the isopyknic chloroform of blended liquid phase, isolated
Chloroform layer, solvent removed by evaporation at reduced pressure, it is dried, obtains extract a, standby；
(5) Rhizoma Polygonati Odorati is pulverized, add 60-70 DEG C of heating and refluxing extraction of methanol according to the ratio that solid-to-liquid ratio is 1g:13-17ml
70-90 minute, filter, obtain filtrate d；
(6) D113 ion exchange resin on filtrate d, filtrate D is 2:1 with the volume ratio of D113 ion exchange resin, tree
The height of fat post and diameter are than for 7:1, and upper column flow rate is 1.4-1.6BV/h, successively with 80% ethanol, 60% second of 2BV of 4BV
Alcohol, 40% ethanol of 1BV, carry out gradient elution, and elution flow rate is 1.4-1.6BV/h, collects 40% ethanol elution；
(7) 40% ethanol elution is evaporated to the extractum without alcohol taste, extract dry, obtains extract b；
(8) extract a and extract b mix homogeneously, Chinese medicine extract is obtained.
HGH and IGF-1 liposome complex capsule, is by HGH and IGF-1 liposome and pharmaceutically acceptable other are auxiliary
Material, makes according to conventional method pharmaceutically.
Present invention have the advantage that
1, HGH and IGF-1 makes liposome according to certain method, further makes capsule, makes HGH and IGF-1 bag
It is enclosed in liposome, it is easier to absorption of human body, overcomes HGH and IGF-1 by injection system just absorbent shortcoming.
2, according to the characteristic of HGH and IGF-1, liposome preparation process is respectively prepared 2 kinds of immobilized artificial membranes, remixes and prepare sky
White liposome turbid liquor, improves envelop rate and the release amount of medicine of HGH and IGF-1 complex liposome, substantially increases drug effect.
3, raw material uses several Chinese medicine, then obtain extract with specific extracting method, many with HGH, IGF-1 and Semen sojae atricolor
Peptide synergism, prepared medicine is possible not only to the growth promotion that keeps healthy, and can improve immunity, blood sugar lowering, has treatment children's
The effect of paralysis.
Specific embodiment
Below in conjunction with specific embodiment, the present invention is described further, to help understanding present disclosure.
Embodiment 1:
HGH and IGF-1 liposome, is made up of the raw material of following masses number: HGH15 part, IGF-1 15 parts, soybean polypeptide
14 parts, green tea 5 parts, Fructus Citri Sarcodactylis 5 parts, Herba Taraxaci 2 parts, Rhizoma Polygonati Odorati 2 parts, 30 parts of lecithin, lipositol 30 parts, 50 parts of cholesterol, poly-
Pyrusussuriensis ester 80 20 parts, ethyl acetate 100 parts, isopropanol 100 parts, phosphate buffer 1 80 parts.
HGH and IGF-1 liposome, preparation method is:
(1) cholesterol of lecithin and half formula ratio is dissolved in ethyl acetate, mix homogeneously, removing is evaporated under reduced pressure molten
Agent, obtains immobilized artificial membrane I；
(2) cholesterol of lipositol He second half formula ratio is dissolved in isopropanol, mix homogeneously, is evaporated under reduced pressure and removes
Solvent, obtains immobilized artificial membrane II；
(3) immobilized artificial membrane I and immobilized artificial membrane II being added to the phosphate buffer of half formula ratio, stirring makes immobilized artificial membrane complete
Fully hydrated, make blank liposome suspension；
(4) HGH, IGF-1, soybean polypeptide and Chinese medicine extract are dissolved in the phosphate buffer of second half formula ratio,
Add blank liposome suspension, obtain HGH and IGF-1 liposome turbid liquor；
(5) HGH and IGF-1 liposome turbid liquor is spray-dried, obtains HGH and IGF-1 liposome.
Wherein, the preparation method of Chinese medicine extract is:
(1) green tea is added distilled water according to the ratio that solid-to-liquid ratio is 1g:16ml, soak 2 hours at 80 DEG C, filter,
Filtrate a；
(2) Herba Taraxaci is added to filtrate a, 200W supersound extraction 30 minutes, filter, obtain filtrate b, standby；
(3) Fructus Citri Sarcodactylis is pulverized, adds 75% ethanol ultrasonic extraction 40 minutes according to the ratio that solid-to-liquid ratio is 1g:9ml, filter,
Obtain filtrate c；
(4) filtrate b and filtrate c is merged, obtain mixed liquor, extract with the isopyknic chloroform of blended liquid phase, isolated
Chloroform layer, solvent removed by evaporation at reduced pressure, it is dried, obtains extract a, standby；
(5) Rhizoma Polygonati Odorati is pulverized, adds 60 DEG C of heating and refluxing extraction of methanol 70 minutes according to the ratio that solid-to-liquid ratio is 1g:13ml,
Filter, obtain filtrate d；
(6) D113 ion exchange resin on filtrate d, filtrate D is 2:1 with the volume ratio of D113 ion exchange resin, tree
The height of fat post and diameter are than for 7:1, and upper column flow rate is 1.4BV/h, successively with 80% ethanol of 4BV, 60% ethanol of 2BV,
40% ethanol of 1BV, carries out gradient elution, and elution flow rate is 1.4BV/h, collects 40% ethanol elution；
(7) 40% ethanol elution is evaporated to the extractum without alcohol taste, extract dry, obtains extract b；
(8) extract a and extract b mix homogeneously, Chinese medicine extract is obtained.
HGH and IGF-1 liposome complex capsule, is by HGH and IGF-1 liposome and pharmaceutically acceptable other are auxiliary
Material, makes according to conventional method pharmaceutically.
Embodiment 2:
HGH and IGF-1 liposome, is made up of the raw material of following masses number: HGH20 part, IGF-1 20 parts, soybean polypeptide
16 parts, green tea 7 parts, Fructus Citri Sarcodactylis 7 parts, Herba Taraxaci 3 parts, Rhizoma Polygonati Odorati 2 parts, 35 parts of lecithin, lipositol 35 parts, 55 parts of cholesterol, poly-
Pyrusussuriensis ester 80 25 parts, ethyl acetate 120 parts, isopropanol 120 parts, phosphate buffer 200 parts.
HGH and IGF-1 liposome, preparation method is:
(1) cholesterol of lecithin and half formula ratio is dissolved in ethyl acetate, mix homogeneously, removing is evaporated under reduced pressure molten
Agent, obtains immobilized artificial membrane I；
(2) cholesterol of lipositol He second half formula ratio is dissolved in isopropanol, mix homogeneously, is evaporated under reduced pressure and removes
Solvent, obtains immobilized artificial membrane II；
(3) immobilized artificial membrane I and immobilized artificial membrane II being added to the phosphate buffer of half formula ratio, stirring makes immobilized artificial membrane complete
Fully hydrated, make blank liposome suspension；
(4) HGH, IGF-1, soybean polypeptide and Chinese medicine extract are dissolved in the phosphate buffer of second half formula ratio,
Add blank liposome suspension, obtain HGH and IGF-1 liposome turbid liquor；
(5) HGH and IGF-1 liposome turbid liquor is spray-dried, obtains HGH and IGF-1 liposome.
Wherein, the preparation method of Chinese medicine extract is:
(1) green tea is added distilled water according to the ratio that solid-to-liquid ratio is 1g:18ml, soaks 2.5 hours at 85 DEG C, filter,
Obtain filtrate a；
(2) Herba Taraxaci is added to filtrate a, 200W supersound extraction 35 minutes, filter, obtain filtrate b, standby；
(3) Fructus Citri Sarcodactylis is pulverized, adds 75% ethanol ultrasonic extraction 45 minutes according to the ratio that solid-to-liquid ratio is 1g:10ml, filter,
Obtain filtrate c；
(4) filtrate b and filtrate c is merged, obtain mixed liquor, extract with the isopyknic chloroform of blended liquid phase, isolated
Chloroform layer, solvent removed by evaporation at reduced pressure, it is dried, obtains extract a, standby；
(5) Rhizoma Polygonati Odorati is pulverized, adds 65 DEG C of heating and refluxing extraction of methanol 80 minutes according to the ratio that solid-to-liquid ratio is 1g:15ml,
Filter, obtain filtrate d；
(6) D113 ion exchange resin on filtrate d, filtrate D is 2:1 with the volume ratio of D113 ion exchange resin, tree
The height of fat post and diameter are than for 7:1, and upper column flow rate is 1.5BV/h, successively with 80% ethanol of 4BV, 60% ethanol of 2BV,
40% ethanol of 1BV, carries out gradient elution, and elution flow rate is 1.5BV/h, collects 40% ethanol elution；
(7) 40% ethanol elution is evaporated to the extractum without alcohol taste, extract dry, obtains extract b；
(8) extract a and extract b mix homogeneously, Chinese medicine extract is obtained.
HGH and IGF-1 liposome complex capsule, is by HGH and IGF-1 liposome and pharmaceutically acceptable other are auxiliary
Material, makes according to conventional method pharmaceutically.
Embodiment 3:
HGH and IGF-1 liposome, is made up of the raw material of following masses number: HGH25 part, IGF-1 25 parts, soybean polypeptide
18 parts, green tea 10 parts, Fructus Citri Sarcodactylis 10 parts, Herba Taraxaci 4 parts, Rhizoma Polygonati Odorati 4 parts, 40 parts of lecithin, lipositol 40 parts, 60 parts of cholesterol,
Polyoxyethylene sorbitan monoleate 30 parts, ethyl acetate 140 parts, isopropanol 140 parts, phosphate buffer 220 parts.
HGH and IGF-1 liposome, preparation method is:
(1) cholesterol of lecithin and half formula ratio is dissolved in ethyl acetate, mix homogeneously, removing is evaporated under reduced pressure molten
Agent, obtains immobilized artificial membrane I；
(2) cholesterol of lipositol He second half formula ratio is dissolved in isopropanol, mix homogeneously, is evaporated under reduced pressure and removes
Solvent, obtains immobilized artificial membrane II；
(3) immobilized artificial membrane I and immobilized artificial membrane II being added to the phosphate buffer of half formula ratio, stirring makes immobilized artificial membrane complete
Fully hydrated, make blank liposome suspension；
(4) HGH, IGF-1, soybean polypeptide and Chinese medicine extract are dissolved in the phosphate buffer of second half formula ratio,
Add blank liposome suspension, obtain HGH and IGF-1 liposome turbid liquor；
(5) HGH and IGF-1 liposome turbid liquor is spray-dried, obtains HGH and IGF-1 liposome.
Wherein, the preparation method of Chinese medicine extract is:
(1) green tea is added distilled water according to the ratio that solid-to-liquid ratio is 1g:20ml, soak 3 hours at 90 DEG C, filter,
Filtrate a；
(2) Herba Taraxaci is added to filtrate a, 200W supersound extraction 40 minutes, filter, obtain filtrate b, standby；
(3) Fructus Citri Sarcodactylis is pulverized, adds 75% ethanol ultrasonic extraction 50 minutes according to the ratio that solid-to-liquid ratio is 1g:11ml, filter,
Obtain filtrate c；
(4) filtrate b and filtrate c is merged, obtain mixed liquor, extract with the isopyknic chloroform of blended liquid phase, isolated
Chloroform layer, solvent removed by evaporation at reduced pressure, it is dried, obtains extract a, standby；
(5) Rhizoma Polygonati Odorati is pulverized, adds 70 DEG C of heating and refluxing extraction of methanol 90 minutes according to the ratio that solid-to-liquid ratio is 1g:17ml,
Filter, obtain filtrate d；
(6) D113 ion exchange resin on filtrate d, filtrate D is 2:1 with the volume ratio of D113 ion exchange resin, tree
The height of fat post and diameter are than for 7:1, and upper column flow rate is 1.6BV/h, successively with 80% ethanol of 4BV, 60% ethanol of 2BV,
40% ethanol of 1BV, carries out gradient elution, and elution flow rate is 1.6BV/h, collects 40% ethanol elution；
(7) 40% ethanol elution is evaporated to the extractum without alcohol taste, extract dry, obtains extract b；
(8) extract a and extract b mix homogeneously, Chinese medicine extract is obtained.
HGH and IGF-1 liposome complex capsule, is by HGH and IGF-1 liposome and pharmaceutically acceptable other are auxiliary
Material, makes according to conventional method pharmaceutically.
Embodiment 4:
HGH and IGF-1 liposome, is made up of the raw material of following masses number: HGH28 part, IGF-1 13 parts, soybean polypeptide
20 parts, green tea 3 parts, Fructus Citri Sarcodactylis 15 parts, Herba Taraxaci 1 part, Rhizoma Polygonati Odorati 5 parts, 25 parts of lecithin, lipositol 45 parts, 45 parts of cholesterol, poly-
Pyrusussuriensis ester 8015 parts, ethyl acetate 150 parts, isopropanol 90 parts, phosphate buffer 230 parts.
Remaining is with embodiment 1.
Embodiment 5:
HGH and IGF-1 liposome, is made up of the raw material of following masses number: HGH20 part, IGF-1 20 parts, soybean polypeptide
16 parts, green tea 7 parts, Herba Taraxaci 3 parts, Rhizoma Polygonati Odorati 3 parts, 35 parts of lecithin, lipositol 35 parts, 55 parts of cholesterol, polyoxyethylene sorbitan monoleate
25 parts, ethyl acetate 120 parts, isopropanol 120 parts, phosphate buffer 200 parts.
Wherein, the preparation method of Chinese medicine extract is:
(1) green tea is added distilled water according to the ratio that solid-to-liquid ratio is 1g:18ml, soaks 2.5 hours at 85 DEG C, filter,
Obtain filtrate a；
(2) Herba Taraxaci is added to filtrate a, 200W supersound extraction 35 minutes, filter, obtain filtrate b, be evaporated to former
The 1/3 of volume, is dried, obtains extract a, standby；
(3) Rhizoma Polygonati Odorati is pulverized, adds 65 DEG C of heating and refluxing extraction of methanol 80 minutes according to the ratio that solid-to-liquid ratio is 1g:15ml,
Filter, obtain filtrate d；
(4) D113 ion exchange resin on filtrate d, filtrate D is 2:1 with the volume ratio of D113 ion exchange resin, tree
The height of fat post and diameter are than for 7:1, and upper column flow rate is 1.5BV/h, successively with 80% ethanol of 4BV, 60% ethanol of 2BV,
40% ethanol of 1BV, carries out gradient elution, and elution flow rate is 1.5BV/h, collects 40% ethanol elution；
(5) 40% ethanol elution is evaporated to the extractum without alcohol taste, extract dry, obtains extract b；
(8) extract a and extract b mix homogeneously, Chinese medicine extract is obtained.
HGH and IGF-1 liposome complex capsule, is by HGH and IGF-1 liposome and pharmaceutically acceptable other are auxiliary
Material, makes according to conventional method pharmaceutically.
Remaining is with embodiment 2.
Embodiment 6:
HGH and IGF-1 liposome, is made up of the raw material of following masses number: HGH20 part, IGF-1 20 parts, soybean polypeptide
16 parts, green tea 7 parts, Fructus Citri Sarcodactylis 7 parts, Herba Taraxaci 3 parts, Rhizoma Polygonati Odorati 3 parts, 35 parts of lecithin, 55 parts of cholesterol, polyoxyethylene sorbitan monoleate 25 parts,
Ethyl acetate 120 parts, phosphate buffer 200 parts.
HGH and IGF-1 liposome, preparation method is:
(1) lecithin and cholesterol are dissolved in ethyl acetate, mix homogeneously, solvent removed by evaporation at reduced pressure, obtain phospholipid
Film；
(2) immobilized artificial membrane being added to the phosphate buffer of half formula ratio, stirring makes the complete aquation of immobilized artificial membrane, makes
Blank liposome suspension；
(3) HGH, IGF-1, soybean polypeptide and Chinese medicine extract are dissolved in the phosphate buffer of second half formula ratio,
Add blank liposome suspension, obtain HGH and IGF-1 liposome turbid liquor；
(4) HGH and IGF-1 liposome turbid liquor is spray-dried, obtains HGH and IGF-1 liposome.
Remaining is with embodiment 2.
Embodiment 7:
HGH and IGF-1 liposome,
Wherein, the preparation method of Chinese medicine extract is:
(1) green tea is added distilled water according to the ratio that solid-to-liquid ratio is 1g:20ml, soak 3 hours at 90 DEG C, filter,
Filtrate a；
(2) Fructus Citri Sarcodactylis is added to filtrate a, 200W supersound extraction 40 minutes, filter, obtain filtrate b, standby；
(3) Herba Taraxaci is pulverized, add 95% ethanol ultrasonic extraction 50 minutes according to the ratio that solid-to-liquid ratio is 1g:11ml, mistake
Filter, obtains filtrate c；
(4) filtrate b and filtrate c is merged, obtain mixed liquor, extract by the isopyknic ethyl acetate of blended liquid phase, separate
Obtain ethyl acetate layer, solvent removed by evaporation at reduced pressure, be dried, obtain extract a, standby；
(5) Rhizoma Polygonati Odorati is pulverized, adds 70 DEG C of heating and refluxing extraction of methanol 90 minutes according to the ratio that solid-to-liquid ratio is 1g:17ml,
Filter, obtain filtrate d；
(6) HD-8 ion exchange resin on filtrate d, filtrate D is 2:1 with the volume ratio of HD-8 ion exchange resin, tree
The height of fat post and diameter are than for 7:1, and upper column flow rate is 1.6BV/h, successively with 80% ethanol of 4BV, 60% ethanol of 2BV,
40% ethanol of 1BV, carries out gradient elution, and elution flow rate is 1.6BV/h, collects 40% ethanol elution；
(7) 40% ethanol elution is evaporated to the extractum without alcohol taste, extract dry, obtains extract b；
(8) extract a and extract b mix homogeneously, Chinese medicine extract is obtained.
Remaining is with embodiment 3.
Embodiment 8
By the arbitrary prepared liposome of embodiment 1-7, according to the operational approach of conventional capsule, add pharmaceutic adjuvant, will
Each component mix homogeneously, pelletizes, and uses conventional capsule to prepare method filling and makes capsule, and the content of the most each component is as follows:
Remaining is with any one in embodiment 1-7.
Embodiment 9
By the arbitrary prepared liposome of embodiment 1-7, according to the operational approach of conventional capsule, add pharmaceutic adjuvant, will
Each component mix homogeneously, pelletizes, and uses conventional capsule to prepare method filling and makes capsule, and the content of the most each component is as follows:
Remaining is with any one in embodiment 1-7.
Embodiment 10
By the arbitrary prepared liposome of embodiment 1-7, according to the operational approach of conventional capsule, add pharmaceutic adjuvant, will
Each component mix homogeneously, pelletizes, and uses conventional capsule to prepare method filling and makes capsule, and the content of the most each component is as follows:
Remaining is with any one in embodiment 1-7.
1, the liposome physicochemical property that the present invention prepares
The drug quality of the liposome that each embodiment prepares is as shown in table 1, and the computational methods of envelop rate are: envelop rate=
(HGH1-HGH2)/HGH1× 100%, HGH1The total amount of the recombinant human somatropin for putting in step (4), HGH2For step (4)
The total amount of the recombinant human somatropin in middle centrifuged supernatant.
The liposome physicochemical property that table 1 present invention prepares
As can be seen from Table 1, in the range of present invention process, the envelop rate of the liposome that (embodiment 1-3) is prepared can
To reach more than 95%, in 30 days, release amount of medicine can reach more than 98%, hence it is evident that higher than embodiment 4, (proportioning raw materials is not
Together), embodiment 5 (few a few herbs raw material), embodiment 6 (a kind of immobilized artificial membrane of preparation), the embodiment 7 (extraction of Chinese medicine extract
Method is different), especially with the best results of embodiment 2, it is seen that present invention HGH, IGF-1 and soybean polypeptide are as the most former
Material, and the combination of green tea, Chinese medicine, use 2 kinds of immobilized artificial membranes to make liposome, specific Traditional Chinese medicine extraction method, prepared liposome
Envelop rate and in 30 days release amount of medicine higher, drug effect and bioavailability are high simultaneously.
2, the present invention effect to improving immunity
Turnover of Mouse Peritoneal Macrophages phagocytosis chicken red blood cell is used to test (half intracorporal method):
Taking 80 mices, body weight 18～24g, be randomly divided into 8 groups, often group 10, is divided into matched group, embodiment 1-7 group, real
Executing the complex liposome 0.2g/10g.BW gavage that example 1-7 group is prepared respectively by embodiment 1-7 group, matched group gives equivalent distillation
Water, once a day, gives 30 days continuously, every Mus lumbar injection 20% chicken erythrocyte suspension 1ml, is spaced 3min, at cervical dislocation
Dead animal, is faced upward position and is fixed on Mus plate, and abdominal skin is cut off in center, rotates Mus plate through abdominal cavity saline injection 2ml.
1min, sucking-off abdominal cavity washing liquid 1ml, average mark drips on 2 microscope slides, puts into the enamel tray being lined with wet gauze, incubates in 37 DEG C
Case incubation 30min, incubates complete, and normal saline rinses, and dries, fixes with 1:1 acetone methanol solution, and 4%Giemsa dyes 3min.Meter
The macrophage number counting phagocytosis chicken red blood cell and the macrophage number swallowed, result is as shown in table 2.
Table 2 present invention shadow noon (means standard deviation) to Turnover of Mouse Peritoneal Macrophages phagocytosis chicken red blood cell test
| Group | Phagocytic percentage (%) | Phagocytic index |
| Matched group | 19.5±3.7 | 0.24±0.06 |
| Embodiment 1 | 41.2±5.1 | 0.52±0.13 |
| Embodiment 2 | 45.7±5.8 | 0.57±0.14 |
| Embodiment 3 | 43.3±5.3 | 0.55±0.10 |
| Embodiment 4 | 36.0±4.5 | 0.43±0.09 |
| Embodiment 5 | 28.1±4.2 | 0.38±0.06 |
| Embodiment 6 | 26.6±4.9 | 0.36±0.08 |
| Embodiment 7 | 27.9±4.0 | 0.35±0.05 |
As can be seen from Table 2, the liposome that (embodiment 1-3) is prepared in the range of present invention process is to mouse peritoneal
What macrophage phagocytic chicken red blood cell was tested is substantially better than embodiment 4 (proportioning raw materials is different), embodiment 5 shadow noon (in few several tastes
Medicine raw material), embodiment 6 (a kind of immobilized artificial membrane of preparation), embodiment 7 (extracting method of Chinese medicine extract is different), especially with reality
Execute the best results of example 2, it is seen that the present invention with HGH, IGF-1 and soybean polypeptide as primary raw material, and green tea, Chinese medicine combination,
Using 2 kinds of immobilized artificial membranes to make liposome, specific Traditional Chinese medicine extraction method, prepared liposome is improved Turnover of Mouse Peritoneal Macrophages
The effect of phagocytosis chicken red blood cell, has potentiation to body immunity.
3, the present invention is to hypoglycemic effect
Choose healthy mice, body weight 30 ± 3g, randomly draw 10 mices as Normal group, remaining mice fasting
Press 200mg/kg lumbar injection 2%ALX after 12h, measure blood glucose after 72h, filter out fasting glucose the mice of 11.1mmol/L is
Diabetic mice.Hyperglycemia mice is randomly divided into model group, embodiment 1-7 group, often group 10, and embodiment 1-7 group is respectively by real
Execute the complex liposome 0.2g/10g.BW gavage that example 1-7 group prepares, the distilled water of the quality such as model group gavage, take one every day
Secondary, continuous 2 weeks, last took front 12h fasting, and after last takes 2h, eye socket takes blood, measured blood sugar content, as shown in table 3.
Table 3 present invention impact (means standard deviation) on blood glucose in diabetic mice
| Group | Blood glucose (mmol/L) |
| Normal group | 5.46±0.82 |
| Model group | 15.17±1.63 |
| Embodiment 1 | 6.28±1.16 |
| Embodiment 2 | 5.82±1.04 |
| Embodiment 3 | 5.95±0.98 |
| Embodiment 4 | 9.63±1.51 |
| Embodiment 5 | 10.50±1.45 |
| Embodiment 6 | 11.64±1.37 |
| Embodiment 7 | 10.86±1.22 |
As can be seen from Table 3, the liposome that (embodiment 1-3) is prepared in the range of present invention process can reduce sugar
The blood glucose of the sick mice of urine, effect is substantially better than embodiment 4 (proportioning raw materials is different), embodiment 5 (few a few herbs raw material), implements
Example 6 (a kind of immobilized artificial membrane of preparation), embodiment 7 (extracting method of Chinese medicine extract is different), especially with the effect of embodiment 2
Good, it is seen that the present invention with HGH, IGF-1 and soybean polypeptide as primary raw material, and green tea, Chinese medicine combination, use 2 kinds of immobilized artificial membranes
Making liposome, specific Traditional Chinese medicine extraction method, prepared liposome has hypoglycemic effect.
4, the present invention effect to treatment poliomyelitis
Select poliomyelitis sequela patient 160 example clinical observation, the maleest 83 examples, women 77 example, age 2-40 year, flat
All 19.6 years old ages, course of disease 1-10, average 4.8 years；Patient is randomly divided into 8 groups, for matched group, embodiment 1-7 group, often organizes 20
Example, often group conditions of patients qualifications and record of service are basically identical, without significant difference, have comparability, and each group is kept on a diet the most on request.
The capsule that the liposome embodiment 8 that embodiment 1-7 group is prepared by embodiment 1-7 group respectively prepares is taken,
Oral, every day twice, each 1, within 30 days, it is a course for the treatment of, takes 3 to 4 courses for the treatment of continuously, the person's of being in a bad way meal with wine of following the doctor's advice adds
Amount；Matched group uses safflower oil, requires to use in strict accordance with description.Result is as shown in table 4.
Outcome measure:
(1) cure: the symptoms such as nerve, muscle, skeleton atrophy gradually take a turn for the better, and finally fully recover.
(2) take a turn for the better: above-mentioned symptom has been alleviated.
(3) invalid: symptom is unchanged.
Table 4 present invention effect to treatment poliomyelitis
As can be seen from Table 4, the liposome complex capsule pair that (embodiment 1-3) is prepared in the range of present invention process
Poliomyelitis has an obvious therapeutic effect, and be better than embodiment 4 (proportioning raw materials is different), (few a few herbs are former for embodiment 5
Material), embodiment 6 (a kind of immobilized artificial membrane of preparation), embodiment 7 (extracting method of Chinese medicine extract is different), especially with embodiment 2
Best results, it is seen that the present invention with HGH, IGF-1 and soybean polypeptide as primary raw material, and green tea, Chinese medicine combination, use 2
Planting immobilized artificial membrane and make liposome, specific Traditional Chinese medicine extraction method, prepared liposome complex capsule has treatment poliomyelitis
Effect.
Claims (4)
1.HGH and IGF-1 liposome, is characterized by: be made up of the raw material of following masses number: HGH15-25 part, IGF-1 15-
25 parts, soybean polypeptide 14-18 part, green tea 5-10 part, Fructus Citri Sarcodactylis 5-10 part, Herba Taraxaci 2-4 part, Rhizoma Polygonati Odorati 2-4 part, lecithin 30-40
Part, lipositol 30-40 part, cholesterol 50-60 part, polyoxyethylene sorbitan monoleate 20-30 part, ethyl acetate 100-140 part, isopropanol
100-140 part, phosphate buffer 1 80-220 part.
2. HGH and IGF-1 liposome as claimed in claim 1, is characterized by: preparation method is:
(1) cholesterol of lecithin and half formula ratio is dissolved in ethyl acetate, mix homogeneously, solvent removed by evaporation at reduced pressure,
Obtain immobilized artificial membrane I；
(2) cholesterol of lipositol He second half formula ratio is dissolved in isopropanol, mix homogeneously, removing is evaporated under reduced pressure molten
Agent, obtains immobilized artificial membrane II；
(3) immobilized artificial membrane I and immobilized artificial membrane II being added to the phosphate buffer of half formula ratio, stirring makes the complete water of immobilized artificial membrane
Change, make blank liposome suspension；
(4) HGH, IGF-1, soybean polypeptide and Chinese medicine extract are dissolved in the phosphate buffer of second half formula ratio, then add
Enter blank liposome suspension, obtain HGH and IGF-1 liposome turbid liquor；
(5) HGH and IGF-1 liposome turbid liquor is spray-dried, obtains HGH and IGF-1 liposome.
3. HGH and IGF-1 liposome as claimed in claim 2, is characterized by: the preparation method of Chinese medicine extract is:
(1) green tea is added distilled water according to the ratio that solid-to-liquid ratio is 1g:16-20ml, soak 2-3 hour at 80-90 DEG C, mistake
Filter, obtains filtrate a；
(2) Herba Taraxaci is added to filtrate a, 200W supersound extraction 30-40 minute, filter, obtain filtrate b, standby；
(3) Fructus Citri Sarcodactylis is pulverized, add 75% ethanol ultrasonic extraction 40-50 minute according to the ratio that solid-to-liquid ratio is 1g:9-11ml, mistake
Filter, obtains filtrate c；
(4) filtrate b and filtrate c is merged, obtain mixed liquor, extract with the isopyknic chloroform of blended liquid phase, isolated chloroform
Layer, solvent removed by evaporation at reduced pressure, it is dried, obtains extract a, standby；
(5) Rhizoma Polygonati Odorati is pulverized, add 60-70 DEG C of heating and refluxing extraction 70-90 of methanol according to the ratio that solid-to-liquid ratio is 1g:13-17ml
Minute, filter, obtain filtrate d；
(6) D113 ion exchange resin on filtrate d, filtrate D is 2:1 with the volume ratio of D113 ion exchange resin, resin column
Height and diameter than for 7:1, upper column flow rate is 1.4-1.6BV/h, successively with 80% ethanol of 4BV, 60% ethanol of 2BV,
40% ethanol of 1BV, carries out gradient elution, and elution flow rate is 1.4-1.6BV/h, collects 40% ethanol elution；
(7) 40% ethanol elution is evaporated to the extractum without alcohol taste, extract dry, obtains extract b；
(8) extract a and extract b mix homogeneously, Chinese medicine extract is obtained.
4.HGH and IGF-1 liposome complex capsule, is characterized by: be HGH and the IGF-1 liposome that prepared by claim 2 and
Other adjuvants pharmaceutically acceptable, make according to conventional method pharmaceutically.
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