CN103070849A - Application of oxyresveratrol in preparing ovary aging resistant medicine - Google Patents
Application of oxyresveratrol in preparing ovary aging resistant medicine Download PDFInfo
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- CN103070849A CN103070849A CN2013100327428A CN201310032742A CN103070849A CN 103070849 A CN103070849 A CN 103070849A CN 2013100327428 A CN2013100327428 A CN 2013100327428A CN 201310032742 A CN201310032742 A CN 201310032742A CN 103070849 A CN103070849 A CN 103070849A
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Abstract
The invention discloses an application of oxyresveratrol in preparing an ovary aging resistant medicine. The oxyresveratrol has significant treatment effects on various ovary aging diseases such as polycystic ovarian syndrome, dysfunctional uterine bleeding, premature ovarian failure and climacteric syndrome. The oxyresveratrol is prepared into the oral, transdermal or injection medicine for treating the ovary aging diseases.
Description
Technical field
The present invention relates to the medical usage of oxidized resveratrol, i.e. the application of oxidized resveratrol in effectively the multiple ovary for the treatment of fails disease medicament.
Background technology
The bioprocess of the complexity that the ovary aging is a multifactor interaction, build up.Constantly consume along with day in year passing follicle, it is the main cause of ovary aging that Follicle number descends; And body intracellular metabolite product piles up that microenvironment changes in the ovary, such as the damage to follicle of free radical, advanced glycation end products, and the continuous accumulation of the factors such as Mitochondrial DNA Mutation, telomere shortening, the quality of residue follicle also reduces simultaneously; The expression of series of genes and changing function in the ovary, and the function of the secretion precision control ovary of H-P-O axle (Hypothalamus-Pituitary-Ovary Axis, hypothalamic-pituitary-ovarian axis) hormone.The ovarian function decline is that ovary is old and feeble, and take menopause as performance, in the past menopause in 40 years old is called premature ovarian failure (Premature Ovarian Failure, POF), and namely the pathologic ovary is old and feeble.The ovarian function decline can cause ovarian dysfunction disease (such as polycystic ovarian syndrome, dysfunctional uterine bleeding etc.), then depletion causes perimenopausal syndrome, each tract senile disease appears in these diseases in succession: coronary heart disease, cerebrovascular incidence rate obviously improve in the blood circulation, easily send out presenile dementia, depression among the central nervous system, osteoporosis is occurred frequently etc. because calcium loss sharply increases.In recent years, along with rhythm of life accelerate and learning pressure excessive, occur in the existing obviously trend of increase of ovary decline disease association disease among the women.Because there are youth, middle age or elderly woman in each family, so the old and feeble above-mentioned disease that produces of ovary become the social problem that generally is concerned about, the research of this disease become focus and the difficult point of domestic and international reproductive development circle.
At present both at home and abroad research thinks that ovary aging and heredity, immunity, metabolism, viral infection and the factor such as iatrogenic are relevant more.Be summarised as the following aspects: 1 karyotype is unusual, by the organon phase to the old and feeble patient of chromosomal abnormality ovary, some syndrome genes mainly concentrate on X chromosome and the autosome, what are relevant with Inheritance for karyotype unusual, and the gene mutation that causes of environmental factors also is the key factor that causes this disease in addition.2 immunity factors, may with cytokine induction granular cell and lutein cell, mainly be histocompatibility complex (MHC) class antigen presentation, bring out the autoimmune response of body, it is relevant that granular cell and follicle are damaged.In addition, think that in addition ovarian function fails and the generation of ovary autoantibody and the autoimmune disease that merges other endocrine bodies of gland or system, relevant such as autoimmune thyroiditis, systemic lupus erythematosus (sle), myasthenia gravis, parathyroid function decline, rheumatoid arthritis, idiopathic thrombocytopenic purpura, diabetes etc.3 promoting sexual gland hormone and receptor abnormality thereof, the conduction defect of promoting sexual gland hormone FSH, LH and receptor thereof (FSHR, LHR) can cause the ovarian function decline.4 azymias can cause galactosemia, and galactosemia occurs relevant with the ovarian function decline.Galactose can directly damage oocyte, and its metabolite can produce infringement to ovary essence again, and the infiltration of galactose molecule also can change the activity of promoting sexual gland hormone, thereby causes the exhaustion of folliculus ovarii.5 ovary destructive factors, day by day frequent along with women's social activity, being not intended to the Long Term Contact reflected ray stimulates, or accepts lonizing radiation heavy dose of or over a long time because of work, disease, contingency, all can destroy ovary.6 other dangerous factors, smoking, depression, the exogenous E of long-term a large amount of contacts
2With nullipara and ovary decline close relationship is arranged.And heavy metal class environmental hormone is still blank on the impact research of ovarian function decline, and along with China's rapid development of economy, this class environmental factors will be grown health to the Chinese people people's livelihood can produce long-range adverse effect.
Hormone replacement therapy (Hormone replacement therapy, HRT) be the important method of the present old and feeble related symptoms of modern medicine treatment ovary and disease, but existing research shows that obviously it has multiple side effect and carcinogenecity at a specified future date, is subject to significant restrictions its clinical practice.Corresponding this state, follow Traditional Chinese medical theory in China, advocate " the kidney invigorating tonifying YIN, soothing the liver nourishing blood " and delay the ovary aging, the treatment of employing Chinese medicinal formulae, but also do not getting clear in the situation of its cause of disease, pathology at present, exist the mistaken ideas of abusing medicine, disorderly treating, and it is unclear that the compound medicine treatment has the effective substance basis, the defective that mechanism of action is not clear can't make Chinese medicine move towards modernization, moves towards internationalization.
Oxidized resveratrol (Oxyresveratrol), have another name called 4-[(E)-2-(3, the 5-dihydroxy phenyl) vinyl] benzene-1, the 3-glycol, belong to the trans stilbene compounds, in various plants, found at present its existence, belong to (Artocarpus) such as the Fructus Artocarpi Heterophylli in the Moraceae (Moraceae), Morus (Moraceae), orange Morus (Maclura), the smilax (Smilax) of Liliaceae (Liliaceae), the Gnetum (Gnetum) of Veratrum (Veraturm) and Gnetaceae (Gnetaceae) etc.Oxidized resveratrol is yellow solid, molecular formula: C
14H
12O
4Molecular weight: 244.24; 199～204 ℃ of fusing points; its structure is suc as formula I; it is a kind of effective tyrosinase inhibitor, in addition also has very strong skin pigment precipitation, and herpes, antiviral, antiinflammatory, antioxidation, neuroprotective and the effect of inhibited apoptosis when cerebral ischemia occurs; owing to have hypotoxicity and water solublity preferably, be fit to carry out drug development.Report oxidized resveratrol for the preparation of the application in the old and feeble medicine of anti-ovary but not yet have at present.
Summary of the invention
Technical problem to be solved by this invention provides the application of oxidized resveratrol in the old and feeble medicine of the anti-ovary of preparation.Described oxidized resveratrol (Oxyresveratrol) has following structure:
The present invention relates to the application of the anti-ovary diseases of aging of oxidized resveratrol (Oxyresveratrol), particularly diseases such as polycystic ovarian syndrome, dysfunctional uterine bleeding, premature ovarian failure or menopausal syndrome etc. are had significant curative effect, but and when being used for such use, have characteristics such as the safety of use and long-term taking.Therefore, oxidized resveratrol can be used in the old and feeble medicine of the anti-ovary of preparation, and this medicine has the purposes of the multiple ovary decline for the treatment of disease.
Oxidized resveratrol can derive from commercially available or adopt prior art to prepare.
The oxidized resveratrol that the present invention uses can be made medicine with any one adjuvant or the drug excipient that pharmaceutically allow, its preparation can be any one dosage form that pharmaceutically allows, and includes but not limited to liquid preparation, granule, tablet, electuary, soft capsule, soft capsule, drop pill, ointment or injection.
Medicine provided by the invention, its form of medication mainly comprises oral administration, percutaneous dosing or drug administration by injection.
The dosage of oxidized resveratrol among the present invention, because of patient's state, body weight, administering mode etc. different different.It is characterized in that described medicine contains oxidized resveratrol and the pharmaceutical carrier of 5-200mg.
Beneficial effect of the present invention compared with the prior art: oxidized resveratrol has significant proliferation function to the gonad granulocyte that exsomatizes, can significantly improve the estrogen secretion level of the gonad granulocyte that exsomatizes, can significantly increase the estrogenic secretion of naturally-aged rat, anti-apoptotic effect to gonad granulocyte is obvious, can reduce Model of Polycystic Ovarian Syndrome Mus FSH, LH, T level, can the sex hormone regulation effect be arranged to caused by cyclophosphamide premature ovarian failure rat.Therefore, oxidized resveratrol can be used in the old and feeble medicine of the anti-ovary of preparation, particularly uses in the medicine that the anti-ovary senile symptom of preparation is premature ovarian failure and resistant ovary syndrome.The present invention is the white Herba chenopodii of oxidation
Reed alcohol provides novel clinical use, has enlarged its range of application.
Description of drawings
Fig. 1 is the apoptotic index figure after the effect of flow cytometer detection of drugs.
The specific embodiment
Below with reference to specific embodiment the present invention is further elaborated, these examples only are used for illustration purpose, and are not used in the restriction scope of the invention.Laboratory animal of the present invention, available from Test Animal Centre, Academy of Military Medical Sciences, P.L.A (production licence number: SCXK (army) 2007-004), selecting clinically has the estradiol valerate of good therapeutic effect to do positive control.Oxidized resveratrol is pressed document (Fu Daxu, Chen Lei, Hou Aijun, Yao Qing, Zhang Wenyu. the chemical constitution study of black Mulberry. Chinese herbal medicine, 2005,36 (9): methods Laboratory self-control 1296-1299), purity 〉=98% (HPLC), the water-soluble administration concentration that is mixed with.
[embodiment one] oxidized resveratrol is to the proliferation function (mtt assay) of the gonad granulocyte that exsomatizes
Get 22-27 age in days female rats, the cervical vertebra dislocation method is put to death the aseptic bilateral ovaries of winning rat behind the conventional 48h of raising, granular cell is discharged the ovary fragmentation, contain with 2mL in the DMEM-F12 culture fluid of penicillin and streptomycin and dilute, and gently piping and druming, make it be dispersed into individual cells.Centrifugal filtration collecting granules cell.With the finite concentration cell suspension inoculation in 24 orifice plates, every hole 80 μ l, every hole adds 120 μ l culture fluid again and adjusts cell density to 200 μ l/ hole, establishes 6 parallel holes for every group.Be placed on 5%CO
2Cultivate 24h in the incubator.Get CO
2Cultivated the granular cell suspension of 24h in the incubator, every hole successively adds modeling concentration Caddy (Cleary) and variable concentrations oxidized resveratrol sample (1.0mgmL
-1, 0.1mgmL
-1, 0.01mgmL
-1), parallel setting Normal group (normal saline) and model group (Caddy (Cleary) and normal saline).In every hole, add MTT solution (5mgmL behind the cellar culture 20h
-1) 20 μ L continuation cultivation 4h, every hole adds three liquid of 80 μ L, and incubated overnight is selected the 570nm wavelength, measures each hole absorbance at enzyme-linked immunosorbent assay instrument, the results are shown in Table 1.Stripped gonad granulocyte cultivation effect all had significant difference (all p＜0.01) with model group after the result showed positive drug, the effect of various dose oxidized resveratrol.
Table 1 oxidized resveratrol is to the proliferation function of the gonad granulocyte that exsomatizes
*P＜0.05,
*P＜0.01vs model group.
ΔP＜0.05,
The Δ ΔP＜0.01vs Normal group
[embodiment two] oxidized resveratrol is to the estrogen and progestogen secretory action (radioimmunology) of the gonad granulocyte that exsomatizes
Get 22-27 age in days female rats, the cervical vertebra dislocation method is put to death the aseptic bilateral ovaries of winning rat behind the conventional 48h of raising, granular cell is discharged the ovary fragmentation, contain with 2ml in the DMEM-F12 culture fluid of penicillin and streptomycin and dilute, and gently piping and druming, make it be dispersed into individual cells.Centrifugal filtration collecting granules cell.With the finite concentration cell suspension inoculation in 24 orifice plates, every hole 80 μ l, every hole adds 120 μ l culture fluid again and adjusts cell density to 200 μ l/ hole, establishes 6 parallel holes for every group.Be placed on 5%CO
2Cultivate 24h in the incubator.Get CO
2Cultivated the granular cell suspension of 24h in the incubator, every hole successively adds modeling concentration Caddy (Cleary) and variable concentrations oxidized resveratrol sample, parallel setting Normal group (normal saline) and model group (Caddy (Cleary) and normal saline).Be placed on 5%CO
2Cultivate 24h in the incubator.Its cell conditioned medium liquid of centrifuging and taking is measured E with radioimmunological kit
2, P content, the results are shown in Table 2.Display model group and Normal group all have significant difference (p＜0.01) as a result, the gonad granulocyte estrogen secretion level that exsomatizes after positive drug, the effect of various dose oxidized resveratrol and model group have significant difference (p＜0.01), and the gonad granulocyte Progesterone Secretion level that exsomatizes after the effect of various dose oxidized resveratrol and model group do not have significant difference.
Table 2 oxidized resveratrol is to the estrogen and progestogen secretory action of the gonad granulocyte that exsomatizes
*P＜0.05,
*P＜0.01vs model group.
ΔP＜0.05,
The Δ ΔP＜0.01vs Normal group
[embodiment three] oxidized resveratrol is on the impact (radioimmunology) of naturally-aged rat estrogen and progestogen secretion
Adopt the senilism model of naturally-aged.Choose 14 month female SD rats, body weight (270 ± 30) g, after Animal adaptability is fed a week, make the colpocytology smear, 4 oestrous cycles of Continuous Observation, with the colpocytology performance oestrous cycle prolong, afterwards Constant Estrus, pseudopregnancy cell phase time repeatedly, as female aged model success.The naturally-aged rat is divided into model group, oxidized resveratrol (3mg/kg, 6mg/kg, 12mg/kg) group and positive drug group at random, every group each 10, other gets 11 of 4 month female rats and is Normal group, every day, gavage was 1 time, Normal group and model group give the isometric(al) normal saline, behind the successive administration 30 days, get blood from rat eye socket venous plexus before the administration and after the last administration, centrifugal, get serum, measure respectively the content of estrogen and progesterone in the serum with radioimmunology, the results are shown in Table 3.Display model group and Normal group all have significant difference (p＜0.01) as a result, gavage positive drug after 30 days estrogen and progestogen all with model group significant difference (p＜0.01) is arranged, and oxidized resveratrol (high, normal, basic dosage) promotes estrogen secretion after 30 days and model group has significant difference (p＜0.01), though the Progesterone Secretion ability is improved, with the model group there was no significant difference.
The comparison of table 3 oxidized resveratrol group and matched group estrogen and progestogen level
*P＜0.05,
*P＜0.01vs model group.
ΔP＜0.05,
The Δ ΔP＜0.01vs Normal group
[embodiment four] oxidized resveratrol is on the impact (flow cytometry) of naturally-aged Cultured Granulosa apoptosis
Adopt the first old model of naturally-aged.Choose 14 month female SD rats, body weight (270 ± 30) g, after Animal adaptability is fed a week, make the colpocytology smear, 4 oestrous cycles of Continuous Observation, with the colpocytology performance oestrous cycle prolong, afterwards Constant Estrus, pseudopregnancy cell phase time repeatedly, as female aged model success.The naturally-aged rat is divided into model group at random, oxidized resveratrol (3mg/kg, 6mg/kg, 12mg/kg) group and positive drug group, every group each 10, other gets 11 of 4 month female rats and is Normal group, every day, gavage was 1 time, Normal group and model group give the isometric(al) normal saline, behind the successive administration 30 days, collect gonad granulocyte after the last administration, carry out flow cytometer and detect (seeing Fig. 1), analyze the relation with apoptosis of administration and gonad granulocyte, the result shows: among the apoptosis figure, UL represents dead cell, and UR represents the apoptosis in late period, and LL represents living cells, LR represents early apoptosis, and UR+LR is the apoptosis total value.Figure 1A (Normal group) apoptosis value is 25.70, Figure 1B (model group) apoptosis value is 86.65, Fig. 1 C (positive controls) apoptosis value is 47.04, Fig. 1 D (oxidized resveratrol high dose group) apoptosis value is 29.78, Fig. 1 E (dosage group in the oxidized resveratrol) apoptosis value is 46.05, Fig. 1 F (oxidized resveratrol low dose group) apoptosis value is 55.43, according to the visible positive drug of numerical value, oxidized resveratrol (high, normal, basic dosage) after 30 days the anti-apoptotic effect to gonad granulocyte obvious.
[embodiment five] oxidized resveratrol is on the impact of Model of Polycystic Ovarian Syndrome rat
24d age teenage SD female rats, body weight 30-40g, 0.5mg18-methylnorethindron day 1 subcutaneous injection, at 27d during age, add with HCG1.5IU day 2 subcutaneous injections, inject altogether 21d.After the modeling success, be divided at random model group, sample (high, normal, basic dosage) group and positive drug group (metformin 0.018g/d), every group each 10, other gets 10 of 45d SD in age female rats and is Normal group, every day, gavage was 1 time, and Normal group and model group give the isometric(al) normal saline, and successive administration is after 15 days, get blood examination after the last administration and survey FSH, LH, T concentration in the serum, the results are shown in Table 4.Display model group and Normal group all have significant difference (p＜0.01) as a result, gavage positive drug, oxidized resveratrol (high, normal, basic dosage) after 15 days FSH, LH, T level the difference of different significances is all arranged from model group.
The comparison of FSH, LH, T concentration in table 4 oxidized resveratrol group and the matched group serum
*P＜0.05,
*P＜0.01vs model group.
ΔP＜0.05,
The Δ ΔP＜0.01vs Normal group
[embodiment six] oxidized resveratrol is on the impact of premature ovarian failure sex hormone level in rats
Adopt 9d SD in age female rats, continuously 14 days cyclophosphamide 8mg/ (kgd) of lumbar injection modeling.After the modeling success, be divided at random model group, sample (high, normal, basic dosage) group and positive drug group (premarin 0.075mg/kg), every group each 10, other gets 10 of 23d SD in age female rats and is Normal group, every day, gavage was 1 time, Normal group and model group give the isometric(al) normal saline, and successive administration was got blood examination and surveyed FSH, E in the serum after 35 days after the last administration
2Concentration the results are shown in Table 5.Display model group and Normal group all have significant difference (p＜0.01) as a result, gavage positive drug, oxidized resveratrol (senior middle school's dosage) FSH, E after 35 days
2Level all has the difference of different significances from model group.
FSH, E in table 5 oxidized resveratrol group and the matched group serum
2The comparison of concentration
*P＜0.05,
*P＜0.01vs model group.
ΔP＜0.05,
The Δ ΔP＜0.01vs Normal group
[embodiment seven] oxidized resveratrol capsule
Oxidized resveratrol 100g, starch 80g, carboxymethyl starch sodium 24g, dextrin 17g, 70% appropriate amount of ethanol is made 1000 (every contains oxidized resveratrol 100mg).
[embodiment eight] oxidized resveratrol capsule
Oxidized resveratrol 15g, starch 10g, carboxymethyl starch sodium 4g, dextrin 12g, 70% appropriate amount of ethanol is made 1000 (every contains oxidized resveratrol 15mg).
[embodiment nine] oxidized resveratrol sheet
Oxidized resveratrol 200g, lactose 55g, amylum pregelatinisatum 15g, hyprolose 10g, magnesium stearate 5g, 70% appropriate amount of ethanol is made 1000 (every contains oxidized resveratrol 200mg).
[embodiment ten] oxidized resveratrol granule
Oxidized resveratrol 6g, lactose powder 800g, magnesium stearate 194g, 70% appropriate amount of ethanol is made 1000 gram granules (containing oxidized resveratrol 6mg in every g granule system).
[embodiment 11] oxidized resveratrol soft capsule
Oxidized resveratrol 5g, soybean oil 10g, gelatin 5g, glycerol 5g, distilled water 7g makes 1000 (every contains oxidized resveratrol 5mg).
[embodiment 12] oxidized resveratrol emulsifiable paste
Oxidized resveratrol 0.5g, stearic acid 100g, hexadecanol 20g, glyceryl monostearate 10g, white oil 10g, methyl hydroxybenzoate 0.8g, butyl hydroxybenzoate 0.2g, glycerol 140g, potassium hydroxide 5g, ethanol 10g, distilled water adds to 1000g.
[embodiment 13] oxidized resveratrol emulsifiable paste
Oxidized resveratrol 0.5g, stearic acid 100g, hexadecanol 20g, glyceryl monostearate 10g, white oil 10g, methyl hydroxybenzoate 0.8g, butyl hydroxybenzoate 0.2g, glycerol 140g, potassium hydroxide 5g, ethanol 10g, distilled water adds to 1000g.
[embodiment 14] oxidized resveratrol gel
Oxidized resveratrol 0.5g, PEG400450g, hexanediol 80g, Tween 80 20g, carbomer 30g, ethanolamine 6ml, benzyl alcohol 10g, distilled water adds to 1000g.
[embodiment 15] oxidized resveratrol injection
Oxidized resveratrol 25.0g is dissolved in the 1000ml water for injection, fully stirring and dissolving, add water for injection to 2000ml, add pin activated carbon 2.5g, be heated to 60 ℃ and stirred 30 minutes, carbon-point filters, filtrate is through 0.25 μ m filtering with microporous membrane degerming, be sub-packed in 1000 cillin bottles, loading amount 2.0ml/ props up, sealing, sterilization, and get final product.
Claims (5)
1. the application of oxidized resveratrol in the old and feeble medicine of the anti-ovary of preparation.
2. according to claim 1 application is characterized in that described ovary senile symptom is polycystic ovarian syndrome, dysfunctional uterine bleeding, premature ovarian failure or menopausal syndrome.
3. according to claim 1 described application is characterized in that described medicine contains oxidized resveratrol and the pharmaceutical carrier of 5-200mg.
4. according to claim 1 described application, the dosage form that it is characterized in that described medicine is any dosage form of pharmaceutically approving.
5. according to claim 4 described application is characterized in that the dosage form of described medicine comprises granule, tablet, electuary, soft capsule, soft capsule, drop pill, ointment or injection.
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| CN110923198A (en) * | 2018-09-19 | 2020-03-27 | 中国农业大学 | Application of resveratrol, in-vitro activating reagent and method of primordial follicles |
| CN113677358A (en) * | 2019-01-19 | 2021-11-19 | 伊斯盖普治疗公司 | Tyrosine inhibitors with immunosuppressive activity in human neonatal keratinocyte progenitor cells |
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