CN1118281C - Application of seeweed polyose sulphate - Google Patents
Application of seeweed polyose sulphate Download PDFInfo
- Publication number
- CN1118281C CN1118281C CN 00120874 CN00120874A CN1118281C CN 1118281 C CN1118281 C CN 1118281C CN 00120874 CN00120874 CN 00120874 CN 00120874 A CN00120874 A CN 00120874A CN 1118281 C CN1118281 C CN 1118281C
- Authority
- CN
- China
- Prior art keywords
- sps
- seaweeds
- sulfated polysaccharides
- purposes according
- polyose
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 title abstract description 5
- 229910021653 sulphate ion Inorganic materials 0.000 title abstract 4
- 208000002672 hepatitis B Diseases 0.000 claims abstract description 13
- 229920001282 polysaccharide Polymers 0.000 claims description 28
- 239000005017 polysaccharide Substances 0.000 claims description 28
- 150000004676 glycans Chemical class 0.000 claims description 27
- 239000003814 drug Substances 0.000 claims description 20
- 239000002775 capsule Substances 0.000 claims description 6
- 238000002347 injection Methods 0.000 claims description 4
- 239000007924 injection Substances 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 claims description 2
- PNNNRSAQSRJVSB-SLPGGIOYSA-N Fucose Natural products C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O PNNNRSAQSRJVSB-SLPGGIOYSA-N 0.000 claims description 2
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical compound C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 claims description 2
- 241000199919 Phaeophyceae Species 0.000 claims description 2
- 230000003203 everyday effect Effects 0.000 claims description 2
- 239000006187 pill Substances 0.000 claims description 2
- 241001474374 Blennius Species 0.000 abstract description 6
- 230000000311 effect on hepatitis Effects 0.000 abstract 1
- 238000012360 testing method Methods 0.000 description 14
- 230000000694 effects Effects 0.000 description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 10
- 229940079593 drug Drugs 0.000 description 10
- 241000700721 Hepatitis B virus Species 0.000 description 8
- 241000699666 Mus <mouse, genus> Species 0.000 description 8
- 241000700605 Viruses Species 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- 241000195474 Sargassum Species 0.000 description 7
- 241000725618 Duck hepatitis B virus Species 0.000 description 6
- 241000272522 Anas Species 0.000 description 5
- 241000272525 Anas platyrhynchos Species 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 208000007271 Substance Withdrawal Syndrome Diseases 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 239000008280 blood Substances 0.000 description 4
- 208000015181 infectious disease Diseases 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 238000000746 purification Methods 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 239000002904 solvent Substances 0.000 description 4
- 230000001629 suppression Effects 0.000 description 4
- 101710142246 External core antigen Proteins 0.000 description 3
- 241000725303 Human immunodeficiency virus Species 0.000 description 3
- 229960004150 aciclovir Drugs 0.000 description 3
- MKUXAQIIEYXACX-UHFFFAOYSA-N aciclovir Chemical compound N1C(N)=NC(=O)C2=C1N(COCCO)C=N2 MKUXAQIIEYXACX-UHFFFAOYSA-N 0.000 description 3
- 230000000840 anti-viral effect Effects 0.000 description 3
- 210000002798 bone marrow cell Anatomy 0.000 description 3
- 150000001720 carbohydrates Chemical class 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 239000012531 culture fluid Substances 0.000 description 3
- 238000002651 drug therapy Methods 0.000 description 3
- 238000007689 inspection Methods 0.000 description 3
- 239000002244 precipitate Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- 208000031404 Chromosome Aberrations Diseases 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- 241000701076 Macacine alphaherpesvirus 1 Species 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 241000607142 Salmonella Species 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 231100000215 acute (single dose) toxicity testing Toxicity 0.000 description 2
- 238000011047 acute toxicity test Methods 0.000 description 2
- 230000017531 blood circulation Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 231100000005 chromosome aberration Toxicity 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 238000009396 hybridization Methods 0.000 description 2
- 210000003141 lower extremity Anatomy 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 210000000653 nervous system Anatomy 0.000 description 2
- 231100000252 nontoxic Toxicity 0.000 description 2
- 230000003000 nontoxic effect Effects 0.000 description 2
- 231100000915 pathological change Toxicity 0.000 description 2
- 230000036285 pathological change Effects 0.000 description 2
- 231100000614 poison Toxicity 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 210000002345 respiratory system Anatomy 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000007790 solid phase Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 2
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 239000003298 DNA probe Substances 0.000 description 1
- 206010013786 Dry skin Diseases 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 241001491705 Macrocystis pyrifera Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical compound CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- MASVCBBIUQRUKL-UHFFFAOYSA-N POPOP Chemical compound C=1N=C(C=2C=CC(=CC=2)C=2OC(=CN=2)C=2C=CC=CC=2)OC=1C1=CC=CC=C1 MASVCBBIUQRUKL-UHFFFAOYSA-N 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 208000004880 Polyuria Diseases 0.000 description 1
- 231100000645 Reed–Muench method Toxicity 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 208000031320 Teratogenesis Diseases 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000003916 acid precipitation Methods 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 235000019270 ammonium chloride Nutrition 0.000 description 1
- 230000000507 anthelmentic effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000002155 anti-virotic effect Effects 0.000 description 1
- 239000003443 antiviral agent Substances 0.000 description 1
- 230000036528 appetite Effects 0.000 description 1
- 235000019789 appetite Nutrition 0.000 description 1
- 206010003246 arthritis Diseases 0.000 description 1
- 238000000376 autoradiography Methods 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 210000000748 cardiovascular system Anatomy 0.000 description 1
- 230000005859 cell recognition Effects 0.000 description 1
- 230000007541 cellular toxicity Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 210000002249 digestive system Anatomy 0.000 description 1
- 230000035619 diuresis Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 125000004185 ester group Chemical group 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 239000003365 glass fiber Substances 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000007952 growth promoter Substances 0.000 description 1
- 230000002489 hematologic effect Effects 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 231100000283 hepatitis Toxicity 0.000 description 1
- 230000003118 histopathologic effect Effects 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000008611 intercellular interaction Effects 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 229930027917 kanamycin Natural products 0.000 description 1
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 1
- 229960000318 kanamycin Drugs 0.000 description 1
- 229930182823 kanamycin A Natural products 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 230000002101 lytic effect Effects 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 201000009240 nasopharyngitis Diseases 0.000 description 1
- 231100000028 nontoxic concentration Toxicity 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 230000007096 poisonous effect Effects 0.000 description 1
- -1 polysaccharide sulfate Chemical class 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000035935 pregnancy Effects 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 235000004252 protein component Nutrition 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 238000003127 radioimmunoassay Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000008054 signal transmission Effects 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 229910052717 sulfur Inorganic materials 0.000 description 1
- 239000011593 sulfur Substances 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 210000001550 testis Anatomy 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 231100000816 toxic dose Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000820 toxicity test Toxicity 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
- 238000009834 vaporization Methods 0.000 description 1
- 230000008016 vaporization Effects 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 230000002618 waking effect Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 230000004584 weight gain Effects 0.000 description 1
- 235000019786 weight gain Nutrition 0.000 description 1
- 239000008096 xylene Substances 0.000 description 1
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention discloses the new medical application of seaweed polyose sulphate in treating hepatitis B. 750 mg to 1500 mg of seaweed polyose sulphate is taken per day in three times, namely that 250 mg to 500 mg of seaweed polyose sulphate is taken every time. The present invention has obvious curative effect on hepatitis B.
Description
The present invention relates to a kind of novel medical use of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS, promptly as the purposes for the treatment of hepatitis B medicament.
Many viral diseases still serious threat human life security, and wherein the hepatitis B (HB) that causes of hepatitis B virus (HBV) is the strong and great infectious disease of hazardness of a kind of infectiousness, does not still have effective medicine at present.
China is the sick big country of sending out of hepatitis B (being called for short hepatitis B later on), and the HBVer reaches 1.2 hundred million, and wherein quite a few patient will be converted into liver cirrhosis and hepatocarcinoma, and it has a strong impact on the healthy of people.The diagnosis and the preventive measure of hepatitis B are mature on the whole, but effectively Drug therapy is unsatisfactory.The most frequently used interferon only reaches 40% to eligible patients's effective percentage in the treatment at present.In view of present employed anti-hbv drug exists therapeutic effect incomplete, phenomenons such as toxic side effect and drug withdrawal bounce-back need be sought medicine effective, low toxicity.
Modern medicinal Sargassum reaches over one hundred kind, has that heat-clearing and toxic substances removing, inducing diuresis to remove edema, the kidney invigorating nourish heart, effects such as activating blood circulation to dissipate blood stasis and the anthelmintic that helps digestion.Contain plurality of active ingredients in the Sargassum, wherein, polysaccharide sulfate is to study comparatively deeply, simultaneously also is to possess the composition of practical value most.Sargassum polysaccharides is a kind of glycoconjugates with special construction, contains the proteoglycan of sulfate group.Drug research based on saccharide has great potential, and carbohydrate molecule plays an important role at aspects such as cell recognition, signal transmission, cell-cell interactions, and sugar moieties has determined the function of whole molecule.The saccharide medicine is at infection, and is anticancer, and all there is exploitation and application in various degree aspects such as antiviral and resisting rheumatoid disease arthritis.Conjugated protein component performance in the polysaccharide suppresses virus breeding and kill virus effect, is that multidigit is put antiviral molecular basis.
To the antivirus action of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS, existing both at home and abroad (Zheng Yongtang, Ben Kunlong, Jin Shanwei, the HIV (human immunodeficiency virus)-resistant activity of protein extract in 17 kind of plant, Chinese virusology, 1998 reported; 13:312-24; Witvrouw M and De Clercq E.Sulfated polysaccharides extracted fromsea algae as potential antiviral drugs.Gen Pharmacol, 1997; 29:497-511; Witvrouw M, Este JA, Mateu MQ, et al.Activity of a sulfated polysaccharidesextracted from the red seaweed aghardhiella tenera against humanimmunodeficiency virus and enveloped viruses.Antiviral chemistry ﹠amp; Chemotherapy, 1994; 5:297-303).But, do not see the research report that its anti-hepatitis B virus effect is arranged.
The present inventor finds that under study for action SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS has the effect of beyond thought significant anti-hepatitis B virus.
The object of the present invention is to provide a kind of new medicine for the treatment of hepatitis B.
SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS of the present invention (Sulfated polysaccharides from seaweeds, SPS) be the protein polysaccharide of a kind of sulfur acid ester group of from Brown algae, extracting, main physicochemical characteristics is: soluble in water, dissolubility 〉=30%, white or yellow polycrystal powder, the distinctive fragrance of tool; SO
4 2-Content is 20 ± 5%, and fucose content is 20 ± 5%, protein content 10-18%.
SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS extraction process of the present invention is by the exploitation of the present inventor's independent studies, and it may further comprise the steps:
1. select materials: the light dry seaweed of optional homemade high-quality (also can use Macrocystis pyrifera (L.) Ag.), the main place of production, China north
The marine site, side;
2. soak and cleaning: soak and cleaning the strict time of controlling immersion and cleaning under the room temperature;
3. pulverize;
4. water is carried;
5. low concentration precipitate with ethanol;
6. the high concentration precipitate with ethanol obtains thick product;
7. purification obtains purified product.
This preparation method mild condition is not destroyed the contained various natural nutrition compositions of Sargassum frond, and is simple to operate, improved the Sargassum comprehensive utilization degree, productive rate higher (2%), and cost is lower, is suitable for suitability for industrialized production.
Pharmaceutical composition of the present invention can be made into tablet, pill, injection and capsule (soft, hard), and every dose contains SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS 250mg, and it is oral to follow the doctor's advice.Every day, consumption 750mg～1500mg can divide to give for three times, each 250mg～500mg.
With reference to State Standard of the People's Republic of China GB15193.3-94, GB15193.5-94, GB15193.8-94, GB15193.7-94, GB15193.4-94, GB15193.14-94 and GB15193.13-94, carry out rat acute toxicity test, acute toxicity test in mice, mouse bone marrow cells micronucleus test (male), mouse bone marrow cells micronucleus test (female), mouse sperm deformity test, mouse testis chromosomal aberration test, Salmonella reversion test, the tertogenicity test of rat system and 30 days feeding trials of rat, the result shows large and small Mus LD
50＞21.50g/kg; Capsule experimental group and group of solvents compare, and the mouse Bone marrow cells micronucleus rate does not have significant difference; Capsule experimental group and group of solvents compare, mouse sperm deformity rate there was no significant difference; Capsule experimental group and group of solvents compare, mice primary spermatocyte chromosome aberrations rate, no significant difference; Salmonella reversion test, experimental group return and become clump count and be less than or equal to nature and return and become 2 times of clump counts; In the rat tertogenicity test, experimental group and group of solvents compare, and index differences such as pregnant Mus pregnancy period weight gain and reproduction embryo inspection, tire Mus growth promoter and skeleton deformity inspection are not remarkable; Rat was fed in 30 days, compared clinical examination with the normal feedstuff matched group: internal organs weighing, blood biochemical are learned inspection, hematological examination, histopathologic examination, difference that there are no significant.It is nontoxic that this result shows that SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS belongs to, hereditary-less toxicity and no teratogenesis material.
The oral SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS capsule of rat (4000mg/kg), one day 2 times, serve on 10 days, under waking state, observe its spiritual nervous system, cardiovascular system respiratory system of unifying, all no abnormal performance of result.
Try clothes through this laboratory scientific and technical personnel and trial volunteer worker 30 people, SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS does not all cause bad variation to digestive system, nervous system, respiratory system and blood circulation, and majority are reflected full of vitality, and appetite increases.5 routine physical weakness, the experimenter who easily catches a cold reflect that times of common cold obviously reduces
Describe the present invention in detail with embodiment below, but these embodiment must not be interpreted as the limitation of the present invention that goes up in all senses.
Embodiment 1: the preparation of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS
The 100kg Sargassum through soaking, clean, pulverizing, adds water 800kg, and 90-100 ℃ extracts 3.5h 3 times.Get the supernatant concentrating under reduced pressure to 25L, precipitate under 30% ethanol condition, get supernatant again, reduction vaporization adds in the ethanol of 85L 95% to 10L, filters, and gets thick product.60 ℃ of dryings, purification, 100 orders are pulverized.
Embodiment 2: the effect of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS resisting HBV virus
SPS has anti-preferably HBV effect in vivo, outward.
One, the external anti-HBV effect research of SPS
1.SPS in the 2.2.15 cell culture to the inhibitory action of HBeAg and HBsAg
(1) cell culture: cover with in the culture bottle of 2.2.15 cell and add 0.25% pancreatin (preparation of Hanks liquid), 37 ℃ digested 3 minutes, add the MEM culture fluid and (contain hyclone 10%, 3% glutamine 1%, G418380g/ml, kanamycin 50U/ml) piping and druming, go down to posterity at 1: 3, covered with in 10 days, and added the cell counting count board counting, be mixed with every milliliter of 100,000 cell inoculation Tissue Culture Plates, the every hole 0.2ml of 96 orifice plates, the every hole 1ml of 24 orifice plates, 37 ℃ of 5%CO2 cultivated 24 hours, and cell experimentizes after growing up to monolayer.
(2) medicine pair cell toxicity test: SPS is mixed with 20mg/ml solution with culture fluid, is diluted to 1.25mg/ml with 2 times and adds 96 porocyte culture plates, and same concentration liquid was changed in per 4 days in every concentration 4 holes, established no drug cell matched group.With the observation of cell pathological changes is index, 8 days microscopically observation of cell pathological changes, and destroying fully is 4; 75% is 3; 50% is 2; 25% is 1; Anosisly become 0.Calculate every concentration liquid average cell lesion degree and suppression ratio (%).Press Reed Muench method and calculate the poisonous concentration of half (TC50), maximal non-toxic concentration (TC0).
(3) medicine is to HBeAg, HBsAg inhibition test: 2.2.15 cell 104/ml inoculates 24 porocyte culture plates, every hole 1ml, 37 ℃ of 5%CO2 cultivated 24 hours, add the following 2 times of dilution test medicinal liquids of non-toxic concn, 5 dilution factors are respectively 10,5,2.5,1.25 and 0.625mg/ml, every concentration 3 holes, 37 ℃ of 5%CO2 cultivate, changed the original content medicinal liquid in per 4 days and cultivate, results culture fluid in the time of the 8th day ,-20 ℃ of stored frozen.The solid phase radioimmunoassay box is measured HBsAg and HBeAg.Effect of drugs calculates: calculate cell contrast and every concentration c pm average and standard deviation, P/N value and inhibition percentage rate (%), medium effective concentration (IC50) and selection index (SI).
2.SPS the inhibitory action ultracentrifugation to hepatitis b virus dna polymerase (HBV DNAp) prepares the HBV granule, add employing virus cracking liquid (0.02M Tris-HCl pH7.6,0.05M NaCl, 0.001M EDTA, 0.1%-mercaptoethanol [ME], 0.01%Triton X-100).Ice bath adds 60ul and mixes liquid (0.2M Tris-HCl pH 7.6,0.2M NH4Cl, 0.06M MgCl2,2%NP-40,0.6%-ME, 0.1mM dNTP), (concentration is respectively 201 SPS: 20,4,0.8,0.16,0.032mg/ml), 30ul lytic virus, 1.5Ci 3H-dTTP, 4 ℃ of abundant mixings, move to 45 ℃ of reactions 2.5 hours, after reaction finishes, add 10% ice-cold trichloroacetic acid precipitation DNA of 3ml, sedimentary DNA closes on the plain film of glass fibre, wash film with 5% trichloroacetic acid and 70% ethanol then,, add the 8ml scintillation solution and (contain 0.5%PPO the film oven dry, the xylene solution of 0.015%POPOP) liquid scintillation counter is measured cpm, calculates suppression ratio.
3.SPS to HBsAg and the bonded inhibition experiment of anti--HBs: with concentration be respectively 20,4,0.8,0.16, the SPS of 0.032mg/ml mixes as testing sample with isopyknic Purification of HBsAg, negative if (no Purification of HBsAg) and positive (no SPS) contrast, with HBsAg content in the solid phase radioimmunological kit working sample, calculate medicine to HBsAg and anti--bonded suppression ratio of HBs.
4. experimental result
(1) SPS has inhibitory action to HBV in 2215 cell culture
| Cytotoxicity (mg/ml) | HbeAg | HBsAg |
| IC 50 TC 0 16.32±0.65 10±0 | IC 50(mg/ml) SI 4.88±0.42 3.37±0.321 | IC 50(mg/ml) SI 4.26±1.04 4.0±1.01 |
(2) SPS has inhibitory action ((IC to the HBV archaeal dna polymerase
50=225 μ g/ml)
(3) SPS inhibition HBsAg combines (IC with anti-HBs
50=200 μ g/ml)
Two, anti-HBV effect research in the SPS body
1. duck hepatitis B virus infection: 1 age in days Beijing duck, clear through the positive Sanguis Anas domestica of lower limb shin intravenous injection Shanghai sheldrake DHBV-DNA, every 0.2ml got blood in back 7 days in infection, separation of serum ,-70 ℃ of preservations are to be checked.
2. Drug therapy test: DHBV infect duckling after 7 days random packet carry out the Drug therapy test, every group of 6 ducklings, 3 dosage groups of administration component, the sulphuric acid Sargassum polysaccharides is respectively 25,50 and 100mg/kg group, lumbar injection 1 day 2 times, 10 days, establish virus control group (DHBV), with the physiologic saline for substitute medicine.Positive drug is made positive control with acyclovir 100mg/kg group.The 7th day is (T0) before the medication after infection, and medication the 5th day (T5) after medication the 10th day (T10) and the drug withdrawal the 3rd day (P3), is got blood from duck lower limb shin vein, separation of serum, and-70 ℃ of preservations are to be checked.
3. detection method: get above-mentioned Sanguis Anas domestica to be checked and check film, measure Sanguis Anas domestica clear in the DHBV-DNA level.Press nick translation test kit description method,, and make the clear dot blot hybridization of Sanguis Anas domestica with 32P labelling DHBV-DNA probe, autoradiography diaphragm speckle, measure OD value (optical filter is 490nm) in microplate reader, calculating serum DHBV-DNA density, with hybridization spot OD value as specimen DHBV-DNA level value.Calculate the inhibition % of (T0) before the 3rd day (P3) serum DHBV-DNA of different time (T5, T10) and drug withdrawal after every group of duck medication and the administration, and mapping, respectively organize the clear DHBV-DNA suppression ratio of Sanguis Anas domestica dynamically.
4. experimental result
DHB (DHBV) infected duck hepatitis animal lumbar injection SPS (50,100 μ g/kg), Bid * 10, with the normal saline group is virus control, do the positive drug contrast with acyclovir (ACV), SPS group (100 μ g/kg) compares with the virus control group, can significantly reduce DHBV infected duck serum DHBV-DNA level.Especially do not have a rebound after it should be noted that drug withdrawal.
Claims (8)
1. the application of SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS in preparation treatment hepatitis B medicament.
2. purposes according to claim 1 is characterized in that said SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS is the Brown algae sulfated polysaccharide.
3. purposes according to claim 2 is characterized in that said SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS water solubility more than or equal to 30%, for white or yellow polycrystalline, fragranced is arranged, SO
4 2-Content is 20 ± 5%, and fucose content is 20 ± 5%, and protein content is 10-18%.
4. purposes according to claim 1 and 2 is characterized in that said SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS gives with tablet form.
5. purposes according to claim 1 and 2 is characterized in that said SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS gives with capsule form.
6. purposes according to claim 1 and 2 is characterized in that said SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS gives with pill.
7. purposes according to claim 1 and 2 is characterized in that said SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS gives with the injection form.
8. purposes according to claim 1 and 2 is characterized in that SULFATED POLYSACCHARIDES FROM SEAWEEDS SPS consumption every day is 750mg～1500mg.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00120874 CN1118281C (en) | 2000-08-04 | 2000-08-04 | Application of seeweed polyose sulphate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00120874 CN1118281C (en) | 2000-08-04 | 2000-08-04 | Application of seeweed polyose sulphate |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1279953A CN1279953A (en) | 2001-01-17 |
| CN1118281C true CN1118281C (en) | 2003-08-20 |
Family
ID=4588451
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 00120874 Expired - Fee Related CN1118281C (en) | 2000-08-04 | 2000-08-04 | Application of seeweed polyose sulphate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1118281C (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101361799B (en) * | 2008-09-01 | 2011-07-20 | 北京世纪博康医药科技有限公司 | Composition containing brown alga polysaccharide sulfuric ester and schisandra chinensis and use thereof |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105273093B (en) * | 2014-07-10 | 2018-01-23 | 中国人民解放军军事医学科学院放射与辐射医学研究所 | A kind of preparation method of seaweed polysaccharide sulfate |
| CN105343121B (en) * | 2015-09-28 | 2018-07-06 | 青岛海洋生物医药研究院股份有限公司 | Application of the guluronic acid sulfuric ester in the drug for preparing anti-hepatitis B virus |
| CN107400172A (en) * | 2017-07-20 | 2017-11-28 | 房俊英 | A kind of bioactive substance extracting method |
-
2000
- 2000-08-04 CN CN 00120874 patent/CN1118281C/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101361799B (en) * | 2008-09-01 | 2011-07-20 | 北京世纪博康医药科技有限公司 | Composition containing brown alga polysaccharide sulfuric ester and schisandra chinensis and use thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1279953A (en) | 2001-01-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104510747B (en) | A kind of new medicine use of iridoid glycoside | |
| CN1704085A (en) | Pharmaceutical composition with immunoregulation function and method for preparing the same | |
| CN1118281C (en) | Application of seeweed polyose sulphate | |
| CN102824417A (en) | New method for treating helicobacter pylori related diseases | |
| EP0382551B1 (en) | Prevention and treatment of herpes virus infections | |
| CN1249072C (en) | Preparation method of golden peach glycoside and its new use in medicine | |
| CN1283462A (en) | Application of astragalin A in preparing medicinal composition | |
| EP0121363A2 (en) | Pyrethroid-containing pharmaceutical compositions | |
| CN1135979C (en) | Application of sophocarpine in preparation of medicine for curing coxsackievirus B myocarditis and its preparation method | |
| CN1128622C (en) | Medicinal composition for treating AlDS | |
| CN1304019C (en) | Compound artemisia rupestris capsule | |
| CN108186625B (en) | Application of RamosissimoneA in the preparation of rheumatoid arthritis drugs | |
| CN1261096C (en) | Compound preparation of reveratrol and application | |
| CN120131654B (en) | Application of tetrandrine in preparation of medicines for preventing and treating swine coronavirus | |
| CN100336552C (en) | Medicine for treating lung cancer | |
| CN101011567A (en) | Medicament for treating AIDS and its preparing method | |
| CN1081065C (en) | Compound oral liquid of cactus | |
| CN100571721C (en) | Long stalk Fructus Schisandrae Sphenantherae extract, Preparation Method And The Use | |
| CN1270038A (en) | Fumigant lotion for treating anorectal disease and preparing process thereof | |
| CN1895300A (en) | Kosam extract, its extraction and use | |
| CN102846748A (en) | Tibetan medicine for treating acquired immunodeficiency syndrome | |
| CN101474247A (en) | Application of spatholobus stem extract in preparing medicament for resisting influenza virus and enterovirus | |
| CN101084991A (en) | Tibetan medicine tangut dragonhead extraction and its application in preparing antivirus medicine | |
| CN1164420A (en) | Preparation for reinforced immunomodulator and its production method | |
| RU2143918C1 (en) | Homeopathic drug "krategus-plus multigran" for treatment of patient with arterial hypertension |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C06 | Publication | ||
| PB01 | Publication | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20030820 Termination date: 20110804 |