CN1151797C - Tumor-blood-vessel growth inhibitor and medicinal composition - Google Patents
Tumor-blood-vessel growth inhibitor and medicinal composition Download PDFInfo
- Publication number
- CN1151797C CN1151797C CNB971226644A CN97122664A CN1151797C CN 1151797 C CN1151797 C CN 1151797C CN B971226644 A CNB971226644 A CN B971226644A CN 97122664 A CN97122664 A CN 97122664A CN 1151797 C CN1151797 C CN 1151797C
- Authority
- CN
- China
- Prior art keywords
- tumor
- blood
- shark cartilage
- vessel growth
- cell
- 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
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/56—Materials from animals other than mammals
- A61K35/60—Fish, e.g. seahorses; Fish eggs
-
- 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
- A61K38/19—Cytokines; Lymphokines; Interferons
- A61K38/20—Interleukins [IL]
- A61K38/208—IL-12
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
- A61K47/14—Esters of carboxylic acids, e.g. fatty acid monoglycerides, medium-chain triglycerides, parabens or PEG fatty acid esters
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Engineering & Computer Science (AREA)
- Marine Sciences & Fisheries (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Zoology (AREA)
- Gastroenterology & Hepatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Immunology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Plant Substances (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
It is an object of the present invention to provide a pharmaceutical composition which enables to administer shark cartilage, which is a tumor-angiogenesis inhibitor, as an anticancer agent. The present invention relates to a tumor-angiogenesis inhibitor which comprises finely divided shark cartilage embedded in a fat or oil matrix, the surface of said matrix being further coated with another lipid differing in a melting point from the said fat or oil.
Description
The present invention relates to have the tumor-blood-vessel growth inhibitor of good anti-cancer activity and analgesic activities, the pharmaceutical compositions that comprises this inhibitor and use said composition treatment method for cancer.
Shark is a kind of fish that belongs to Chondrichthyes, haunts in the coastal waters.Its skeleton is a cartilage, knownly tells the shark cartilage that cartilage obtains then and has anti-tumor activity by catching and slaughter shark, and is effective to the treatment cancer.For example, the cartilage products C artilaid that obtains from the America shark goes on the market as antitumorigenic substance.
When growing to about 1-2mm
3During size, tumor produces angiogenesis factor, and like this, tumor can obtain required nutrition and the oxygen of himself growth.Since to this neovascularity inhibition of proliferation help to suppress the growth of tumor cell and in treatment of cancer effectively, therefore, people have in searching and are just carrying out various researchs aspect the material of angiogenesis inhibiting activity, and known shark cartilage has this angiogenesis inhibiting activity.
Yet, suppress the required shark cartilage of angiogenesis every day effective dose up to 50-60g, and its taste and abnormal smells from the patient are offensive, therefore, in many cases, are difficult to oral administration.Because shark cartilage is the mixture of mucopolysaccharide, except that oral route, does not have other suitable route of administration, therefore, when oral administration, it is insoluble that shark cartilage must keep in the stomach under being in strong acidic condition, but is dissolved in the intestinal of absorption site.For very weak cancer patient, oral administration often is difficult to carry out.Do not solve above-mentioned taste, abnormal smells from the patient and Dose Problem, shark cartilage can not be used as can widely used effective cancer-resisting substance.
Therefore, pressing for can be with the actual technology as effective tumor-blood-vessel growth inhibitor of shark cartilage.
At this moment, find that interleukin 12 (IL-12) is one of cytokine that has the effect of NK (NKT) cell-stimulating.Thereafter research discloses, and but the interleukin 12 stimulation has the active T cell of specific cytotoxic (killer T cell) to tumor cell growth and activated T cell, and the interferon gamma (IFN-γ) that it can promote to activate killer T cell generates.Therefore, as the material that can be used for treating the cancer patient, it has caused extensive attention.
Recently in the U.S., available technique for gene engineering (reorganization IL-12 (rt-IL-12)) mass production IL-12.Existing people attempts directly importing in the cancerous cell with will the encode gene of interleukin 12 of gene transfer technique, so that IL-12 can be applied directly on the cancerous cell.
Suppress tumor growth or the method for its disappearance is comprised from the outside to throw in IL-12 by effective use IL-12, and another kind of method comprises organism is induced from body IL-12 to organism.These advantages from body IL-12 are that it does not cause the danger that abnormal immune is replied.In addition, its sensitivity is strong.Therefore, it is expected to produce the effect that makes tumor dwindle or eradicate significantly.
In view of above-mentioned prior art, the objective of the invention is to, provide to make to be the shark cartilage of tumor-blood-vessel growth inhibitor pharmaceutical compositions as the cancer therapy drug administration.Another object of the present invention is to, the very useful anti-cancer composition that has share these tumor-blood-vessel growth inhibitors and IL-12 inducer is provided.
The present invention relates to tumor-blood-vessel growth inhibitor, the particle diameter that this inhibitor comprises the process pulverize at low temperature that is embedded in fat or the oil matrix is no more than the shark cartilage that 32 microns granule accounts for 99.5 weight % at least, and this stromal surface is higher than the further coating of lipid of above-mentioned fat or oil again by another fusing point.
Below the present invention is described in detail.
The shark cartilage of the key component of tumor-blood-vessel growth inhibitor of the present invention must be the form of pulverizing.In this manual, " form of pulverizing " speech is meant that particle diameter is the 1-90 micron.For preparing the shark cartilage of such pulverizing, preferably the shark cartilage of commercially available specification is pulverized.Because protein and other active component meeting thermal denaturation when the temperature heating that surpasses 60 ℃ in the shark cartilage also lose its effect, therefore need pulverize at low temperatures.Pulverizing at low temperatures can be for example by under-196 ℃ with liquid nitrogen with the shark cartilage flash freezing, with cutter-type pulverizer such as Orient stone roller machine etc. frozen block is carried out rough pulverizing then, then the rough thing piece of pulverizing that is in freezing state is pulverized, and the gains piece is further pulverized with jet-propelled pulverizer such as jet mill etc. with high speed rotor formula pulverizer such as Impeller stone roller machine etc.
When preparation tumor-blood-vessel growth inhibitor of the present invention, the shark cartilage of above-mentioned pulverizing is embedded in fat or the oil matrix.
The fat or the oil that constitute this fat or oil matrix are not limited to any particular types, as long as it is generally used for the pharmacy purpose.Therefore, can suitably select this, for example, can be from Oleum Glycines, vegetable oil, Oleum Cocois, Oleum Arachidis hypogaeae semen and Semen Gossypii wet goods hardened vegetable oils; Sclerosis such as Adeps Bovis seu Bubali, Adeps Sus domestica and fish oil animal oil; And select in their mixture.In above-mentioned fat and oil, preferred fusing point is 42 ℃-56 ℃ a fat and oily.If necessary, can share with antioxidant such as vitamin E or catechin etc.
Be embedded in as the requirement of the above-mentioned fat of substrate or the method in the oil not strict to shark cartilage with above-mentioned pulverizing.For example, above-mentioned embedding can be by carrying out with the agitated mixture granulating of high-speed stirred type granulating machine with the shark cartilage of fatty or the oily and pulverizing of liquefaction.
To carry out surface coatings with the fat that contains the pulverizing shark cartilage that is embedded in wherein that aforesaid way produces another lipid different with the fusing point of this fat or oil then with the oil matrix reuse.The lipid that in the present invention practice, uses can be selected from the front operation in be used for preparing the fat of fat or oil matrix or oil different, have more dystectic those lipids usually.Object lesson has waxes such as palm wax, rice wax (rice wax) and Cera Flava; Sclerosis animal oil or vegetable oil; Fatty acid such as stearic acid and Palmic acid; Natural resins such as Lac; With long-chain alcohols such as palmityl alcohol and stearyl alcohols.When carrying out above-mentioned coating, for example can use the powder coating technology.
Another method that can use in preparation tumor-blood-vessel growth inhibitor of the present invention for example comprises following operation: the shark cartilage of above-mentioned pulverizing is dispersed in the fat or oil of fusing in advance; make the dispersion liquid cooling with nebulization, in the rolling granulating machine, use fusing point and above-mentioned fat or another different lipid coating gained powder of oil then.
The tumor-blood-vessel growth inhibitor of the present invention that makes thus contains the shark cartilage as main active, and its taste and abnormal smells from the patient are sheltered fully, is difficult to oral problem thereby thoroughly solved.In addition, the mantle friction of the tumor-blood-vessel growth inhibitor that makes reduces, its result, and its flowability improves, and adhesiveness descends, thereby is fit to very much oral.In addition, it can be designed to insoluble under the strong acidic condition of stomach and dissolve in the intestinal at its absorption site.
In the shark cartilage of the pulverizing of using in the present invention practice, particle diameter is no more than 32 microns particle and accounts for 99.5 weight % at least.As describing in detail in an embodiment, the particle diameter of the shark cartilage of pulverizing has very big influence to effect of the present invention.When particle diameter was too big, tumor-angiogenesis inhibiting activity descended.The method that the sieve of the useful required sieve mesh of example of the method for the shark cartilage of preparation uniform particle diameter sieves the shark cartilage of pulverizing.
Though the shark as the raw material of the shark cartilage of use in the present invention's practice is not limited to any particular types, most preferably the shark cartilage that obtains from blue shark fish (blue shark).The cartilage of blue shark fish contains a large amount of active component protein and copolymerization polysaccharide, and is especially suitable.U.S. products C artilaid makes from the mixture of many shark kinds, the difference between therefore existing batch.Another shortcoming is that its effect is lower.
Confirmed already that tumor-blood-vessel growth inhibitor of the present invention not only had tumor-blood-vessel growth and suppresses active, but also has reliable analgesic activities.Cancer patient, especially patient with advanced cancer generally can be told serious pain.Therefore, when taking for the patient who tells serious pain as anticarcinogen tumor-blood-vessel growth inhibitor of the present invention, its active anticancer and analgesic activities produce synergism, thereby can obtain very effective therapeutic effect.
As describing in detail in the following embodiments, there is apoptosis in result of study prompting to how tumor-blood-vessel growth inhibitor of the present invention produces tumor-angiogenesis inhibiting activity.
Though tumor-blood-vessel growth inhibitor of the present invention is very useful as anticarcinogen, it also similarly has analgesic activities, can be used for other pharmaceutical fields.Therefore, in second aspect, the invention provides the pharmaceutical compositions that comprises tumor-blood-vessel growth inhibitor of the present invention.
And, use tumor-blood-vessel growth inhibitor of the present invention with the dosage that is enough to suppress tumor-blood-vessel growth and treat method for cancer and also fall within the scope of the invention.
Pharmaceutical compositions of the present invention is characterised in that it contains tumor-blood-vessel growth inhibitor and interleukin 12 inducer.This pharmaceutical compositions reflection a second aspect of the present invention.Describe a second aspect of the present invention below in detail.
In this description and claims, " interleukin 12 inducer " speech not only is meant the material that can induce interleukin 12 (IL-12) in vivo, but also comprise cancerous cell, the rt-IL-12 that IL-12 itself, IL-12 gene import and put it briefly, all can induce the material that produces IL-12 in vivo, and do not have any particular determination.
Its example has active hemicellulose chemical compound (AHCC) etc.Known AHCC is for handling the biological active substances that obtains by the Plant fiber in the mushroom mycelium cell wall being carried out enzyme, and it contains β-(1 → 3)-D-glucosan, β-(1 → 6)-D-glucosan and α-heteroglycans such as (1 → 4)-D-glucosan; Peptidoglycan, Dan Baijutang, agglutinin, nucleic acid, indigestible polysaccharide etc.
Except that AHCC, the example of the above-mentioned IL-12 inducer that can use in the present invention also comprises the composition of mushroom mycelium.Described mushroom mycelium composition is not limited to any specific kind, but comprises PSK, the SPG of Schizophyllum commune Franch (Schizophyllum commune) mycelium composition of variable color polypor (Polyporus versicolor) the mycelium composition that uses as anticarcinogen and the lentinan of Lentinus Edodes (Lentinula edodes) mycelium composition.
The example of above-mentioned mushroom mycelium composition also comprises Agaricus (agarics), Ganoderma (Ganodermalucidum), the white pore fungi (Albatrellus confluens) of joining, Agaricus blazei, huge Tricholoma mongolicum Imai bacterium (Tricholoma giganteum), the fine pore fungi (Inonotus obliquus) of askew side, Grifola frondosa, Hericium erinaceus (Bull. Ex Fr.) Pers. (Hericium erinaceum), oyster cap fungus (Pleurotus ostreatus), Panellusserotinus, wood layer hole strain (Phellinus), birch pleat pore fungi (Lenzites betulinus), the mushroom (Tricholoma matsutake) of becoming less severe, Perenniporia fraxinea, the mycelia composition of Flammulina velutipes etc.
The IL-12 inducer that can use in the present invention also comprises the fibre composition of streptococcus pyogenes (Streptococcuspyogenes) or haemolitycus.The example of above-mentioned fibre composition comprises known anticarcinogen, as OK-432 (picibanil).
They are to reply dressing agent (BRM) and known substances as biological.
Above-mentioned IL-12 inducer has activation and starts the effect of killer T cell.Therefore, being present in intravital killer T cell is activated by above-mentioned IL-12 inducer.
On the other hand, the neovascularity that the tumor-blood-vessel growth inhibitor of describing in detail above suppresses to be caused by cancerous cell (tumor cell) is bred.When neovascularity propagation was suppressed, under the repressed pressure of blood supply, tumor cell was at adhesion factors such as its surface expression such as Fas antigen, CD80 and CD86.Killer T cell can discern Fas antigen, CD80, CD86 and other are as antigenic adhesion factor.Therefore, these adhesion factors of being expressed are in the state that can easily be discerned by killer T cell, are modified thus and the tumor cell of the easily identification that becomes is attacked by killer T cell easily.
If be in such condition, compare when only using the IL-12 inducer, above-mentioned IL-12 can attack tumor cell more significantly.The collaborative of tumor-blood-vessel growth inhibitor of the present invention and IL-12 inducer is based on above-mentioned mechanism and the very strong antitumaous effect of performance, thereby causes the ruined apoptosis of tumor cell.This discovery is finished the earliest by the present inventor.
The dosage form of giving the pharmaceutical compositions of the present invention that people or other animals take is not had any special restriction, can be that normally used medicine is carried on any dosage form on the carrier.
As above-mentioned carrier, use at least one in solid, semisolid, liquid diluent, filler and other prescription adjuvants, its ratio is 0.1-99.5% for example, is preferably 0.5-90%.Pharmaceutical compositions of the present invention can oral fully or non-oral administration.The example of non-oral route has topical (as organizing interior injection), subcutaneous injection, intramuscular injection, intra-arterial injection or intravenous injection and rectally.The dosage form that is fit to these route of administration can be made with the technological means of routine.
When with above-mentioned composition during, need to determine dosage according to patient's age and body weight, route of administration, disease and the order of severity thereof and other factors as anticarcinogen for example.But represent for adult's oral dose with the weight of active substance,, be preferably in the scope of 5-6 gram/day generally in 1-10 gram/day.Non-oral dose can be according to route of administration and great changes have taken place, generally can be at 100-10,000 milligram/day, be preferably in the scope of 2-5 gram/day.In some cases, dosage is smaller also enough, and in other cases, then may need bigger dosage.The medicament of every day can be divided and take for 2-4 time.
Can use solid or liquid unit dosage forms, for example use dosage forms such as powder, powder, granule, tablet, capsule, syrup, elixir, suspension.
Powder can make by active substance being ground into suitable fineness.Powder can be by being ground into active substance suitable fineness and with itself and the pharmaceutical carrier that has crushed with same way as, for example edible carbohydrate such as starch or mannitol or some other suitable vehicle are mixed and made.If necessary, can add other additives, as flavoring agent, antiseptic, dispersant, coloring agent, spice etc.
Capsule can be made granule or tablet by the powder or the powder that will obtain as stated above, then gained granule bag is assisted in capsule shells such as gelatine capsule and makes.Outside filling, can add lubricant and/or mobile improving agent as required, as silica sol, Pulvis Talci, magnesium stearate, calcium stearate or solid polyethylene glycol.When adopting capsule, can add disintegrating agent or solubilizing agent, wait and improve drug effect as carboxymethyl cellulose, carboxymethylcellulose calcium, low hydroxypropyl cellulose, cross-linking sodium carboxymethyl cellulose, calcium carbonate, the sodium carbonate that replaces.
Soft capsule can suspend or be dispersed in then the gained dispersion liquid to be wrapped in the gelatin foil in vegetable oil, Polyethylene Glycol, glycerol or the surfactant and make by the active substance that will pulverize.
Granule can be made with following method: the active substance of powder type is mixed with above-mentioned excipient and disintegrating agent, (for example add wetting agent then, syrup, gelatinized corn starch, arabic gum, cellulose solution, polymer solution), if it is necessary, also (for example can add binding agent, sodium carboxymethyl cellulose, hydroxypropyl cellulose, methylcellulose, hydroxypropyl emthylcellulose, gelatin, polyvinyl pyrrolidone, polyvinyl alcohol), then knead the gained mixture and the pressurization this mixture is sieved.Except that above-mentioned powder particle method, also available following method prepares granule: earlier mixture is sent in the pelleter, ground the non-completeness sheet piece of gained then.Can add resistance solvent (as paraffin, wax, hardened castor oil), heavy absorbent (for example, quaternary salt) or adsorbent (for example, Bentonite, Kaolin, dicalcium phosphate) earlier.
Tablet can be by adding stearic acid, stearate, Pulvis Talci, mineral oil and lubricant etc. in the granule that obtains with said method, then the gained mixture is made tablet and make.The plain sheet that available film or sweet tablet make thus.
Also available following method is made tablet: active substance of the present invention is mixed with flowable inert carrier, directly make tablet then and save above-mentioned granulating or form the operation of sheet piece.The coating that also can use the transparent or translucent protective coating of making by air-locked Lac coating, make by sugar or macromolecular material and the polishing coating of making by wax.
Other peroral dosage forms such as syrup, elixir and suspension also can be made into unit dosage forms, so that its specified quantitative contains the active substance of certain specified quantitative.Syrup is by making in the aqueous solution that active substance is dissolved in suitable seasoning.Elixir is made by using avirulent alcohols carrier.Suspension is mixed with by active substance is dispersed in the avirulent carrier.Similarly, can at random add suspending agent or emulsifying agent (for example, ethoxyquin isooctadecanol, polyoxyethylene sorbitol ester), antiseptic, flavoring agent (for example, Oleum menthae, glucide) and/or other materials.
Can as required unit dose medicament for oral use be carried out microcyst.Further the above-mentioned medicament of coating or with polymer or wax embedding active substance can prolong the action time of medicament and can obtain slow release.
By injectable compositions such as the solution or the suspension of preparation liquid unit dosage forms, can carry out subcutaneous, intramuscular, intra-arterial or intravenous administration.These preparations can be dissolved or suspended in avirulent liquid-carrier such as the aqueous that is fit to the injection purpose by the active substance with a certain specified quantitative or contain in the oil solvent, then gained solution or suspension are carried out sterilization treatment and make.In the bottle that the injection of also active substance of the powder of a certain specified quantitative or lyophilized form can being packed into is used, then bottle and content thereof are carried out sterilization treatment and sealing.In this case, can prepare standby bottle and carrier before facing injection, to dissolve or to mix.Can add avirulent salt or imbibitions such as saline solution so that composition for injection.In addition, can add use stabilizing agent, antiseptic, suspending agent, emulsifying agent and/or other materials etc.
Rectally can be by with active substance and hydrophobicity or hydrophilic suppository base such as Polyethylene Glycol, cocoa butter, senior ester (as the Palmic acid myristin etc.) or their mixture mixes and the gained mixture ground make with dosage form.
As mentioned above, tumor-blood-vessel growth inhibitor of the present invention does not have the disgusting taste and the abnormal smells from the patient of shark cartilage, and is acidproof and only dissolve in intestinal, but by oral route conventional method administration.In addition, it has reliable active anticancer and analgesic activities, and is very useful as pharmaceutical compositions.
Tumor-blood-vessel growth inhibitor of the present invention and IL-12 inducer are share, can produce very strong active anticancer, therefore very useful to the preparation pharmaceutical compositions.
Implement best mode of the present invention
Following Production Example and embodiment further describe the present invention by way of example.But they do not limit scope of the present invention.
Production Example 1
Be embedded in the manufacturing of the coated preparation of the shark cartilage in oil or the fat
With 60 weight portion mean diameters is the shark cartilage adding high-speed stirred formula granulating machine (super mixer of 27 microns pulverizing; Dark river Industrial Co., Ltd product) in, the granulating machine internal temperature is remained on 10 ℃ and continue to stir with 20-100rpm in, spray into the fixed oil that derives from Adeps Bovis seu Bubali (fusing point: 48 ℃) of 10 weight portion molten conditions.The droplet of melting oil sticks on the shark cartilage of pulverizing, forms matrix.But in this stage, shark cartilage partly is exposed to the surface, and the coating situation is still insufficient.Therefore; sclerosis vegetable oil (fusing point: 68 ℃) each initial substrate of coating with the pulverizing that to be 5 microns, fusing point different with the fusing point of the above-mentioned fixed oil that derives from Adeps Bovis seu Bubali of 30 weight portion mean diameters; coating method is for to add described sclerosis vegetable oil in the same stirring-type granulating machine; with about 1; the speed of 000rpm stirred the mixture 30 minutes, made described sclerosis vegetable oil attach and be coated with all over the initial substrate surface thus.Thereby obtain very gratifying preparation with enough acid resistances of having of lipid substrate coating and sustained release performance.
Embodiment 1
Test to mouse tumor-angiogenesis inhibiting activity
(the shark cartilage particle diameter is estimated)
Test is carried out with back air bag method.Micropore filter is fixed on diameter 5mm plastic hoop both sides and inject 1 * 10
6MH-134 tumor cell and culture medium make test box thus.Mice is divided into 6 one group, and with the back of subcutaneous each mice of implantation of above-mentioned test box (chamber).Give 4 groups of mices with 1, the following shark cartilage of the oral dose of 000mg/kg reaches 4.The 5th, estimate angiogenesis and suppress degree.In another group (matched group), do not take shark cartilage, with the inhibition degree of same way as evaluation on 5th to mice.
The material of promotion angiogenesis leaks from tumor by micropore filter and causes forming neovascularity on the surface that is contacting with test box.The neovascularity of these tumor inducings is shape in the shape of a spiral.By following benchmark this angiogenesis inhibition degree is scored:
(++) 3 minutes: angiogenesis is remarkable
(+) 2 minutes: angiogenesis is clear
(±) 1 minute: angiogenesis is slight
(-) 0 minute: no blood vessel generates
Suppressing percentage ratio uses the minimizing (%) of comparing score with matched group to represent.
Shark cartilage (1): Cartilaid, the America Neoretna is as the raw material shark cartilage., and grind machine (Orient grinds machine) with cutter-type and pulverize roughly this product flash freezing with liquid nitrogen (196 ℃).Then,, grind machine (Impeller grinds machine) with the high speed rotor formula and will further be ground, use jet-propelled stone roller machine (jet mill) to grind at last by ground product still at freezing state.
Shark cartilage (2):, use its sieve top with the sieve of above-mentioned shark cartilage (1) with 32 microns.
Shark cartilage (3):, use its screened part with the sieve of above-mentioned shark cartilage (1) with 32 microns.
The results are shown in Table 1.
Table 1
| ++ (3 minutes) | + (2 minutes) | ± (1 minute) | -(0 minute) | Score | Suppress % | |
| Matched group | 4 mices | 2 mices | 0 mice | 0 mice | 2.7±0.5 | -- |
| Shark cartilage (1) | 2 mices | 3 mices | 1 mice | 0 mice | 2.2±0.8 | 18.5 |
| Shark cartilage (1) | 2 mices | 4 mices | 0 mice | 0 mice | 2.3±0.5 | 14.8 |
| Shark cartilage (1) | 1 mice | 4 mices | 1 mice | 0 mice | 2.0±0.8 | 25.9 * |
*P＜0.05
The above results shows, in the shark cartilage powder of test, and the shark cartilage of the pulverizing by 32 tm screen
Show obviously higher angiogenesis inhibiting activity.
Embodiment 2
Test to mouse tumor-angiogenesis inhibiting activity
(Cartilaid is to Buttershark)
Cartilaid (America shark cartilage) and the cartilage (Bettershark) that derives from blue shark are compared by the mode identical with back air bag method with embodiment 1.The results are shown in Table 2.
The inhibition % of 100mg/kg Cartilaid group (compare with matched group, score reduces percentage ratio) is 3.6%, and 1,000mg/kg Cartilaid group is 21.4%, has significant difference (P＜0.05).In contrast, the inhibition % of the 100mg/kg of Bettershark group is 17.9%, has significant difference (P＜0.05), 1, and the inhibition % of 000mg/kg group is up to 35.7%, and its difference has the significance (P＜0.002) on the statistics.Also find the significant difference (P＜0.05) between Cartilaid and the Buttershark.
Table 2
| Dosage (mg/kg) | ++ (3 minutes) | + (2 minutes) | ± (1 minute) | -(0 minute) | Score | Suppress % | |
| Matched group | -- | 9 mices | 1 mice | 0 mice | 0 mice | 2.8±0.4 | -- |
| Cartilaid | 1,000 | 3 mices | 6 mices | 1 mice | 0 mice | 2.2±0.8 | 21.4 * |
| Cartilaid | 100 | 6 mices | 4 mices | 0 mice | 0 mice | 2.7±0.5 | 3.6 |
| Bettershark | 1,000 | 3 mices | 2 mices | 5 mices | 0 mice | 1.8±1.0 | 35.7 ** |
| Bettershark | 100 | 5 mices | 4 mices | 1 mice | 0 mice | 2.3±0.8 | 17.9 * |
*P＜0.05，
**P＜0.002
The above results shows that Bettershark has the tumor-angiogenesis inhibiting activity that is higher than Cartilaid.
Embodiment 3
Test (seeking the test of Buttershark effective dose) to mouse tumor-angiogenesis inhibition
With 1 * 10
6Each mice of the subcutaneous implantation of tumor cell (sarcoma 180) (the ICR mice, male, 6 the week ages) the back, totally 3 groups, 10 every group.In a group, give the oral Cartilaid of mice (U.S.'s product) behind the implantation tumour cell immediately, dosage is every day 1,000mg/kg, totally 20 days.In another group, (the particle diameter adjusted is to being no more than 32 microns to take the Bettershark of same dose to mice during identical; Through confirming, when pH2, place and still stablized in 3 hours).In the 3rd group, do not give any medicine to mouse feeding 20 days.After 25 days, mice is all slaughtered, tumor is weighed.Gained the results are shown in Table 3.Suppressing percentage ratio uses the minimizing percentage of comparing tumor weight with matched group recently to represent.In the Cartilaid group, compare with matched group, suppressing percentage ratio is 11.1%, and in the Bettershark group, then is 47.0%, has significant difference (P＜0.001).Comparative result between Cartilaid and the Bettershark is shown that Bettershark has stronger anti-tumor activity (P＜0.05).
Table 3
| Average tumor weight (g) | Suppress % | |
| Matched group | 2.752±1.388 | -- |
| Cartilaid | 2.446±1.139 | 11.1 * |
| Bettershark | 1.457±1.965 | 47.0 ** |
*P＜0.05，
**P＜0.001
The The above results prompting, the Bettershark (particle diameter is no more than 32 microns) of everyone 20 gram dosage will be enough effective in clinical practice every day.
Embodiment 3
Test to the mice analgesic activity
Estimate the analgesic activity of Bettershark with acetic acid tumbling (acetic acid writhing method).ICR mice (male, 5 ages in week) is divided into 5 groups, 10 every group.To take for each group to substances by following plan, after 40 days, give injection 2% acetic acid in each mouse peritoneum.During the 10-20 day behind the injection acetic acid, the reflection of rolling that each mice occurs is counted.Behind the injection acetic acid, take substances to mice and until termination the reflection of rolling is counted every day.
In the Cartilaid group, the oral daily dose of Cartilaid is 1,000mg/kg; In the Bettershark group, the oral daily dose of Bettershark is 1,000mg/kg; In the Indacin group, Indacin (indomethacin, on-steroidal analgesic; Commercially available product) oral dose is 25mg/kg; In the chondroitin sulfate group, the oral dose of chondroitin sulfate (commercially available product) is 333mg/kg; In matched group, do not give any medicine to mouse feeding.Suppressing percentage ratio uses the minimizing percentage ratio of comparing above-mentioned reflection number with matched group to represent.The results are shown in Table 4.Aspect analgesic activity, Bettershark has advantage similarly.
Table 4
| The reflection number rolls | Suppress % | |
| Matched group | 31.2±9.6 | -- |
| Cartilaid | 24.3±3.9 | 23.6 * |
| Bettershark | 21.2±10.4 | 32.1 * |
| Indacin | 10.3±5.6 | 67.0 ** |
| Chondroitin sulfate | 23.4±4.5 | 25.0 * |
*P＜0.05，
**P＜0.001
Embodiment 5
The antigenic expression of immunologic opsonin in taking the clinical case of Bettershark
One 80 years old male tells that anticardium has uncomfortable sensation.According to the inspection of gastroscope, he is diagnosed as at stomach angle (gastric angle) has IIc to carry out sexual type gastric cancer (signet-ring cell carcinoma).Biopsy sample is carried out ultramicroscope and histological examination (immunologic test).Owing to his myocardial infarction, therefore can not undergo surgery.
Take used shark cartilage (3) among the embodiment 1 (below, in embodiment 5, be called " Bettershark ") every day for above-mentioned patient with the daily doses of 20 grams.Electron micrograph of carrying out before taking Bettershark and immunology specific stain check that (gastroscopy for the first time) only provided the general finding of common cancerous cell, shows apoptotic finding seldom.Both do not observe adhesion factor (fluorescent antibody technique) and do not observed Fas antigen (special staining) yet, and, do not observe HSP60 (stress protein, heatshock protein; Fluorescent antibody technique) expression.The gastroscopy of carrying out behind 2 first quarter moons of the gastroscopy first time is covered by normal mucosa around having provided and having shown cancer just significantly, and the cancer protuberance has the finding of the trend that is flattened all around.The electron micrograph result shows in the biopsy second time, and cell membrane and Cytoplasm do not have special variation, but clearly illustrates the contraction and the fragmentation of cancerous cell nuclear, and observes apoptotic bodies (apoptotic bodies).Obtain showing the true finding of cancer cell-apoptosis thus.
Meanwhile, the living tissue sample is carried out the immunology specific stain.With anti-HSP60 antibody, anti-CD80 antibody, anti-CD86 antibody and anti-Fas antibody check cancerous cell.Take the result of Bettershark preparation, Fas antigen becomes the positive, and the adhesion factor of HSP60 antigen and CD80, CD86 also becomes the positive.
In gastroscopy for the third time (in the gastroscopy first time after about 5 months), the cancer complete obiteration at last stomach angle.Take the living tissue sample to check at the position of finding gastric cancer, all do not find cancerous cell.
Above-mentioned finding can be summarized as follows.The adhesion factor of unobservable Fas antigen, HSP60 antigen and CD80, CD86 all becomes the positive after taking 2 first quarter moons of Bettershark preparation before taking the Bettershark preparation.Thereafter, gastroscopy for the third time shows, the cancer complete obiteration.This phenomenon can be made description below.Owing to taken the Bettershark preparation, the blood supply of tumor is suppressed, its result, stress protein (HSP60) is expressed, and the adhesion factor of Fas antigen and CD80, CD86 becomes the positive.In for the third time gastroscopy, confirm tumor fully disappear thereafter.
Immunology result of study to tumor shows that tumor regression comprises necrocytosis and apoptosis.Necrocytosis is the phenomenon that is disappeared by the tumor that inflammation or other factors cause under the effect of nonspecific immunity, and wherein cell membrane, Cytoplasm and nucleus are rotten successively, and tumor cell breaks.Under the situation of necrocytosis, the last appearance of histology wherein has neutrophilic leukocyte, macrophage and fibroblast to infiltrate, and then calcification and generation fiber cause cancer to disappear.Its result leaves a trace at this position.
On the other hand, in the situation that the tumor that is caused by apoptosis disappears, inflammation does not take place at all, does not leave a trace yet, and but finds by normal tissue displacement.In this case, known killer T cell is identified in the Fas antigen of tumor cell surface and adheres to, simultaneously with CD80 or/and the co-induction apoptosis such as adhesion factor of CD86.Nuclear degeneration (nucleorhexis and formation apoptosis body) at first occurs, then Cytoplasm and cell membrane are by macrophage phagocytic.Thereby do not leave a trace.
Take in the situation of Bettershark preparation with other above-mentioned, do not obtain pointing out inflammation vestige such as calcification or fibrogenic finding at the position that cancer disappears, prompting thus, the disappearance of cancer should be because due to the apoptosis.This shows disappear in cancer is during very short (1 month to 1 first quarter moon).
Embodiment 6
Immunologic opsonin test to mice
The sarcoma 180 tumor cell is implanted the back of each mice.Tumor growth inhibitory action of more following each group: one group of peritoneal injection TNP470 (military field chemical industry Co., Ltd. product) 10mg, one group of oral Bettershark 1,000mg/kg, one group of peritoneal injection angiostatin (chemotherapy and serum therapy institute product), 24 μ g, matched group then do not give any medicine.After 2 weeks and after 3 weeks, find to compare with matched group, the tumor growth of all administration groups has significant difference and is suppressed (P＜0.05).In the above-mentioned time immunohistology inspection is carried out in the expression of HSP60, Fas antigen, CD80 and the CD86 of tumor tissues, found that the expression of HSP60, Fas antigen, CD80 and CD86 significantly improves.
In addition, lack mice with the T cell, promptly nude mouse carries out test same as described above.Each medicine all shows tumor growth inhibitory action to a certain degree, but does not have significant difference.According to the result that immunohistology is checked, find the positive that is expressed as of HSP60, Fas antigen, CD80 and CD86.But do not obtain the apoptotic finding of any prompting.
The The above results prompting, the anti-tumor activity of angiogenesis inhibitor is not only based on the pure prevention to the tumor blood supply, be suppressed or the expression of Fas antigen, CD80 and CD86 the identification stress protein occurs under the state of qualitative or quantitative variation and expresses at blood supply but also comprise killer T cell tumor, thereby the anti-tumor activity that shows them, cell death inducing thus.
Embodiment 7-10
And with the situation of AHCC and Bettershark
Take AHCC every day for 4 cancer patients shown in the table 5 with the daily dose of 3.0g, meanwhile, take Bettershark every day for them with the daily dose of 20g.After taking 3 months, estimate therapeutic outcome and determine NK activity, IL-12 level and CD4/CD8 value.In embodiment 7 and 10, tumor all reduces 50% or more.But the NK activity then still is in normal range, active no any significant raising.In embodiment 7 and 10, IL-12 is far above normal range.Therefore, can infer that tumor is dwindled relevant with IL-12.
In table 5, CD4 is the helper T cell sign, and CD8 is the killer T cell sign, the increase degree of CD4/CD8 value reflection killer T cell number.This value increases less than 1 expression killer T cell number, and be 1.0-1.5 normal range.
Table 5
| Embodiment | The patient | Symptom | The NK activity | IL-12 in the serum | Therapeutic outcome | CD4/CD8 |
| 7 | 72 years old male | Carrying out property gastric cancer | 24％ | 240ng/mL | Disappear fully | 0.34 |
| 8 | 57 years old male | Carcinoma of cecum cancerous peritonitis late period | 62％ | 230ng/mL | Disappear fully | 0.42 |
| 9 | 69 years old male | Stomach cancer metastasis is to liver late period | 62％ | 103ng/mL | Disappear fully | 0.62 |
| 10 | 71 years old women | Stomach cancer metastasis is to lung late period | 68％ | 78ng/mL | Part disappears | 0.84 |
Claims (5)
1. tumor-blood-vessel growth inhibitor, the particle diameter that comprises the process pulverize at low temperature that is embedded in fat or the oil matrix is no more than the shark cartilage that 32 microns granule accounts for 99.5 weight % at least, and the surface of this substrate is higher than the further coating of lipid of above-mentioned fat or oil matrix by another fusing point.
2. tumor-blood-vessel growth inhibitor as claimed in claim 1 is characterized in that shark cartilage derives from blue shark.
3. anti-cancer composition, it comprises claim 1 or 2 described tumor-blood-vessel growth inhibitors as basis.
4. anti-cancer composition according to claim 3 is characterized in that it also comprises the interleukin 12 inducer.
5. analgesic composition, it comprises claim 1 or 2 described tumor-blood-vessel growth inhibitors.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP08324595A JP3103513B2 (en) | 1996-11-19 | 1996-11-19 | Shark cartilage oral ingestion preparation |
| JP324595/1996 | 1996-11-19 | ||
| JP324595/96 | 1996-11-19 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1185319A CN1185319A (en) | 1998-06-24 |
| CN1151797C true CN1151797C (en) | 2004-06-02 |
Family
ID=18167579
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB971226644A Expired - Fee Related CN1151797C (en) | 1996-11-19 | 1997-11-17 | Tumor-blood-vessel growth inhibitor and medicinal composition |
Country Status (4)
| Country | Link |
|---|---|
| JP (1) | JP3103513B2 (en) |
| KR (1) | KR100362025B1 (en) |
| CN (1) | CN1151797C (en) |
| TW (1) | TW577745B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1198639C (en) * | 2000-01-05 | 2005-04-27 | 吴羽化学工业株式会社 | Immune enhancing compositions |
| JPWO2003030938A1 (en) * | 2001-10-09 | 2005-01-20 | 株式会社オリエントキャンサーセラピー | Cancer immunotherapy |
| KR100971599B1 (en) | 2002-05-15 | 2010-07-20 | 비에이취엔 가부시끼가이샤 | Composition for Preventing or Treating Blood Vessel-Related Disease |
| AU2003264488A1 (en) * | 2002-09-19 | 2004-04-08 | Orient Cancer Therapy Co., Ltd. | Immunotherapeutic for cancer |
| JPWO2005067974A1 (en) * | 2004-01-19 | 2007-12-27 | 株式会社オリエントキャンサーセラピー | Inhibitors of bone metastasis of cancer |
| CN103798785A (en) * | 2012-11-15 | 2014-05-21 | 李恩泽 | Shark cartilage food with subset energy and capable of inhibiting proliferation of tumor and cancer cells |
-
1996
- 1996-11-19 JP JP08324595A patent/JP3103513B2/en not_active Expired - Lifetime
-
1997
- 1997-11-17 KR KR1019970060573A patent/KR100362025B1/en not_active IP Right Cessation
- 1997-11-17 CN CNB971226644A patent/CN1151797C/en not_active Expired - Fee Related
- 1997-11-19 TW TW086117254A patent/TW577745B/en not_active IP Right Cessation
Also Published As
| Publication number | Publication date |
|---|---|
| JP3103513B2 (en) | 2000-10-30 |
| KR100362025B1 (en) | 2003-08-19 |
| CN1185319A (en) | 1998-06-24 |
| JPH10147534A (en) | 1998-06-02 |
| KR19980042506A (en) | 1998-08-17 |
| TW577745B (en) | 2004-03-01 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP1389466B1 (en) | A dispersion on the basis of beta-glucan containing superfine particles, a corresponding process of manufacturing and the use of said dispersion | |
| Namba | Maitake D-fraction: healing and preventive potential for cancer | |
| US7977379B2 (en) | Method for angiogenesis inhibition or immunostimulation | |
| CN104840481A (en) | Composite fungal polysaccharide product | |
| Van Doan et al. | Mushrooms, seaweed, and their derivatives as functional feed additives for aquaculture: an updated view | |
| EP2397147A1 (en) | Feed for preventing and/or treating diseases due to clostridium sp. bacteria in livestock, and anti-clostridium agent | |
| Hassan et al. | Influence of oyster mushroom waste on growth performance, immunity and intestinal morphology compared with antibiotics in broiler chickens | |
| CN1151797C (en) | Tumor-blood-vessel growth inhibitor and medicinal composition | |
| Savelkoul et al. | Immunomodulatory effects of mushroom β-glucans | |
| Ibuki et al. | Effect of dietary β-1, 4-mannobiose on the growth of growing broiler chicks | |
| CN1526444A (en) | Composition for immunological activation | |
| CN1485072A (en) | Coix seed oil soft capsule for curing prostate diseases and the application thereof | |
| US20010009903A1 (en) | Augmentation method for antitumor activity of substance containing amygdalin, composition containing augmented amygdalin contained substance, method for assessing antitumor efficacy of treatment for amygdalin contained substance and method for assessing antitumor, substance containing amygdalin | |
| WO2009116507A1 (en) | Enteral nutrient | |
| JP4752233B2 (en) | Immunostimulator | |
| JP2005089388A (en) | Agent for enhancing immunopotentiative action | |
| CN102379905A (en) | Ganoderma lucidum spore oil-containing Chinese medicinal composition with efficient cancer resistance | |
| JP2004121070A (en) | Health food | |
| JP2001048795A (en) | Preparation which comprise fine powdery shark cartilage, can be expected to have anti-tumor effect, and can orally be taken | |
| JP2001097881A (en) | Cancer prophylaxis agent, and cancer prophylaxis food or feed all derived from grifola frondosa | |
| CN1960717A (en) | Composition for capsule film, capsule film, and capsule made with the same | |
| CN1850107A (en) | Fatty cream composition for maligant splanchnocoele hydrops and its preparing method | |
| JP3858073B2 (en) | High-functional health food and method for producing high-performance health food | |
| WO2001070251A1 (en) | Anticancer compositions | |
| CN108785323A (en) | The pulullan polysaccharide that spermine as immunopotentiator is modified |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: K. K. KIYOARATA ENTERPRISE Free format text: FORMER OWNER: K. K. KIYOARATA ENTERPRISE BAMERTIANXUBANG Effective date: 20050318 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20050318 Address after: Tokyo, Japan Co-patentee after: K. K. Kiyoarata Enterprise Patentee after: Japan Oil and Grease Ltd. Address before: Tokyo, Japan Co-patentee before: K. K. Kiyoarata Enterprise Patentee before: Japan Oil and Grease Ltd. Co-patentee before: Yagita Akikuni |
|
| REG | Reference to a national code |
Ref country code: HK Ref legal event code: GR Ref document number: 1046294 Country of ref document: HK |
|
| C56 | Change in the name or address of the patentee |
Owner name: JAPAN OIL CO.; K. K. KIYOARATA ENTERPRISE Free format text: FORMER NAME OR ADDRESS: NOF CORP.; K. K. KIYOARATA ENTERPRISE |
|
| CP01 | Change in the name or title of a patent holder |
Address after: Tokyo Co-patentee after: K. K. Kiyoarata Enterprise Patentee after: Japanese oil company Address before: Tokyo Co-patentee before: K. K. Kiyoarata Enterprise Patentee before: NOF Corporation |
|
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20040602 Termination date: 20151117 |
|
| EXPY | Termination of patent right or utility model |