CN110038027A - For inhibiting the medical composition of inflammation - Google Patents
For inhibiting the medical composition of inflammation Download PDFInfo
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- CN110038027A CN110038027A CN201810041014.6A CN201810041014A CN110038027A CN 110038027 A CN110038027 A CN 110038027A CN 201810041014 A CN201810041014 A CN 201810041014A CN 110038027 A CN110038027 A CN 110038027A
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- vitamin
- inflammation
- sodium hyaluronate
- inhibiting
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
- A61K31/353—3,4-Dihydrobenzopyrans, e.g. chroman, catechin
- A61K31/355—Tocopherols, e.g. vitamin E
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/365—Lactones
- A61K31/375—Ascorbic acid, i.e. vitamin C; Salts thereof
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/726—Glycosaminoglycans, i.e. mucopolysaccharides
- A61K31/728—Hyaluronic acid
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Abstract
It is a kind of for inhibiting the medical composition of inflammation, it includes (a) sodium hyaluronate, (b) vitamin and (c) the acceptable carriers of medicine, wherein composition (b) is at least one of vitamin C and vitamin E；And the weight ratio of composition (a) and (b) are 1:0.02 to 1:1.
Description
Technical field
The medical composition more particularly to a kind of medicinal combination of Antiarthritic that the present invention relates to a kind of for inhibiting inflammation
Object.
Background technique
Arthritis is a common chronic disease, causes arthritis because of articular cartilage degeneration or connective tissue inflammation
Bitterly, so influence joint normal activity.According to happening part and reason, arthritis can be divided into 100 multiple types altogether, most often
The person of opinion includes osteoarthritis (degenerative arthritis), rheumatoid arthritis, urarthritis, bacterial arthritis, stiff
Property arthritis vertebralis and erythematosus lupus etc..
For arthritic treatment, conservative, the therapy of Non-surgical, such as drug therapy are usually first taken in initial stage
It is treated with injection, when initial therapy does not become effective, can just use the therapeutic modality of surgical operation.Known drug is treated
Use the anti-inflammatory drug of steroids and on-steroidal, wherein although steroid medicine has analgesic effect fast and significant excellent
Point, but many side effects can be caused, such as osteoporosis, wound are not easy to heal, upper gastrointestinal bleeding, or even can aggravate such as height
The symptom of the diseases such as blood pressure or diabetes, therefore it is not included into the ranks of drug therapy gradually.As for non-steroidal drug, although
Also there is good analgesic effect, if but be used for a long time, such as peptic ulcer, edema of lower extremity and renal function damage can be generated
The side effects such as wound, therefore be restricted in also in practical application more.
Sodium hyaluronate (hyaluronic acid), also known as " hyaluronic acid " or " uronic acid ", it is known that it is imitated with anti-inflammatory
Fruit, therefore be employed for extensively in the injection for inhibiting osteoarthritis.Wherein, extremely by the injection direct injection containing sodium hyaluronate
The feeling of pain of inflammation and sufferer is slowed down in the joint of patient with appropriateness.The mechanism of action of sodium hyaluronate is so far and indefinite, but it is known its
A kind of lubricant is alternatively arranged as to help joint motion, while achieving the effect that improve function of joint.However, those skilled in the art
Although member is it has been found that sodium hyaluronate can effectively mitigate pain, it can cause temporarily within a few days instead sometimes after entering human body
When property inflammatory response, serious person even causes chronic inflammation, and (this can be found in Leopold et al., Increased
frequency of acute local reaction to intra-articular hylan GF-20(Synvisc)in
patients receiving more than one course of treatment.J Bone Joint Surg,2002；
84:1619-23；Bernardeau et al.,Acute arthritis after intra-articular
hyaluronate injection:onset of effusions without crystal.Ann Rheum Dis,2001；
60:518-20；And Kroesen et al., Induction of an acute attack of calcium
pyrophosphate dihydrate arthritis by intra-articular injection of hylan G-F
20(Synvisc).Clin Rheumatol,2000；19:147-9, they's entirety are simultaneously for reference in herein).Therefore,
If the anti-inflammatory effect to be reached can be provided with lower sodium hyaluronate dosage, it can slow down, even sodium hyaluronate be avoided to enter human body
The subsequent inflammatory response that may cause later.
Also a kind of method for improving sodium hyaluronate preparation of reported in literature passes through sodium hyaluronate in the way of chemical synthesis
One victory peptide be connected to on the active aminomethyl folic acid (methotrexate, MTX) of anti-inflammatory to form a conjugate
(conjugate), (this can be found in Homma et al., Novel to anti-inflammatory effect of the thus obtained product with improvement
hyaluronic acid-methotrexate conjugates for osteoarthritis treatment,
Bioorganic and Medicinal Chemistry, 17 (2009), 4647-4656, the document full text and in herein for
With reference to).But according to the teaching of the document, simple mixing sodium hyaluronate is formed by mixture with aminomethyl folic acid and can not
Reach improved effect, to synthesize the means that the conjugate necessitates.However, victory peptide must be used by preparing the conjugate
Raw material and it is related to complicated synthesis step, this certainly will increase the manufacturing cost of sodium hyaluronate preparation, not only in causing on batch production technique
Difficulty also increases the financial burden of user, therefore has limitation in clinical application more.
The inventors discovered that and with sodium hyaluronate and vitamin C and/or vitamin E, it is possible to provide the anti-inflammatory effect of enhancement,
The anti-inflammatory effect to be reached is provided so as to lower sodium hyaluronate dosage, mitigates or exempts sodium hyaluronate and enter institute after human body
The temporary inflammatory response that may cause.
Summary of the invention
A purpose of the present invention is that provide it is a kind of for inhibiting the medical composition of inflammation, it includes: (a) sodium hyaluronate,
(b) vitamin and (c) the acceptable carrier of medicine；Wherein, composition (b) is at least one of vitamin C and vitamin E；And at
The weight ratio of part (a) and (b) are 1:0.02 to 1:1.
Wherein, the average molecular weight of composition (a) is about 30,000 to about 10,000,000 dalton (Dalton).
Wherein, the average molecular weight of composition (a) is about 500,000 to about 6,000,000 dalton.
Wherein, the weight ratio of composition (a) and (b) are 1:0.05 to 1:0.5.
Wherein, the medical composition for inhibiting inflammation is in for oral, subcutaneous injection, intravenous injection or intra-articular note
Penetrate the form of dispensing.
Wherein, described for inhibiting the medical composition of inflammation for Antiarthritic.
Wherein, described for inhibiting the medical composition of inflammation for anti-osteoarthritis, resisting rheumatoid arthritis or resisting
Urarthritis.
Another object of the present invention is to provide a kind of methods for inhibiting inflammation, and it includes so that a patient is applied an effective quantity
Above-mentioned medical composition.
A purpose of the present invention is that providing a kind of for inhibiting the set group of inflammation comprising one includes the glass of effective dose
The second part of the first part of uric acid and the vitamin comprising effective dose；Wherein, the vitamin be vitamin C with
At least one of vitamin E；And the weight ratio of the sodium hyaluronate and the vitamin is 1:0.02 to 1:1.
Wherein, the first part and second part are in the solution form for injection dispensing, and the set group further includes an injection
Syringe.
Detailed description of the invention
Fig. 1, which is shown, to be used alone or and the fibroblast sample synovial cell that is handled with sodium hyaluronate and vitamin C
The statistics column of the performance amount of inflammation medium tumor necrosis factor-alpha (TNF-α) in (fibroblast-like synoviocytes)
Shape figure；
Fig. 2, which is shown, to be used alone or and in the fibroblast sample synovial cell of sodium hyaluronate and vitamin C processing
The statistics column diagram of the performance amount of inflammation medium Jie Bai Su -1 β (IL-1 β)；
Fig. 3, which is shown, to be used alone or and in the fibroblast sample synovial cell of sodium hyaluronate and vitamin E processing
The statistics column diagram of the performance amount of TNF-α；
Fig. 4, which is shown, to be used alone or and in the fibroblast sample synovial cell of sodium hyaluronate and vitamin E processing
The statistics column diagram of the performance amount of inflammation medium IL-1 β；
Fig. 5-1 to 5-4, which is shown, to be used alone or and the fibroblast sample synovial membrane that is handled with sodium hyaluronate and vitamin C
The statistics column diagram of the performance amount of inflammation correlation factor in cell, wherein Fig. 5-1 shows the performance amount of IL-1, Fig. 5-2 display
The performance amount of IL-8, Fig. 5-3 show the performance amount of MMP-3, and Fig. 5-4 shows performance amount (" * " expression p < 0.05 of TNF-α；"**"
Indicate p < 0.01)；
Fig. 6-1 to 6-4, which is shown, to be used alone or and the fibroblast sample synovial membrane that is handled with sodium hyaluronate and vitamin E
The statistics column diagram of the performance amount of inflammation correlation factor in cell, wherein Fig. 6-1 shows the performance amount of IL-1, Fig. 6-2 display
The performance amount of IL-8, Fig. 6-3 show the performance amount of MMP-3, and Fig. 6-4 shows performance amount (" * " expression p < 0.05 of TNF-α；"**"
Indicate p < 0.01)；And
Fig. 7-1 to 7-5 is shown through and with sodium hyaluronate and inflammation in the fibroblast sample synovial cell of vitamin C processing
The statistics column diagram of the performance amount of correlation factor, wherein Fig. 7-1 shows the performance amount of IL-6, and Fig. 7-2 shows the performance of IL-8
Amount, Fig. 7-3 show the performance amount of TNF-α, and Fig. 7-4 shows the performance amount of IL-10, and Fig. 7-5 shows performance amount (" * " table of MMP-3
Show p < 0.05；" * * " indicates p < 0.01).
Specific embodiment
Detailed technology and preferred embodiment of the invention, will be described in the following contents, for fields skill of the present invention
Feature of the invention is illustrated in art personnel accordingly.
In this article (especially after attached claim in), unless otherwise stated, used " one ", "the" and similar
Term is interpreted as comprising odd number and plural form.
As described above, after sodium hyaluronate enters human body, sometimes in causing temporary inflammatory response in a few days, or even cause
Chronic inflammation is unfavorable for the anti-inflammatory benefit to be provided；On the other hand, also there is what document proposed to prepare sodium hyaluronate and amido
The modification method of the conjugate of methopterin, then have many restrictions.The inventors discovered that compared to sodium hyaluronate, dimension is used alone
Raw element C or vitamin E, the present invention and with sodium hyaluronate and vitamin C Ji when ╱ or vitamin E handle cell, it is possible to provide with not
Inhibition inflammation effect of expected multiplication effect (synergistic effect, the i.e. effect of 1+1 > 2).The present inventor also sends out
It is existing, compared to and with sodium hyaluronate and relatively small amount vitamin C, and with sodium hyaluronate and relatively great amount of ascorbic anti-inflammatory
Effect is obviously poor, shows and can not provide the anti-Guan Fayan with synergistic effect with sodium hyaluronate and relatively great amount of vitamin C
Effect.The present invention can simply means be improved now by sodium hyaluronate and the vitamin C of relatively small amount and/or being used in combination for vitamin E
The defect of some sodium hyaluronate preparations.
Therefore, the present invention provides a kind of for inhibiting the medical composition of inflammation, it includes: (a) sodium hyaluronate, the life of (b) dimension
The acceptable carrier of element and (c) medicine；Wherein, composition (b) is at least one of vitamin C and vitamin E；And composition (a) with
(b) weight ratio is 1:0.02 to 1:1.
Sodium hyaluronate is one of the main component for constituting extracellular matrix, is widely distributed in endothelial tissue, connective tissue, epidermal tissue
The physiological activities such as hyperplasia and migration and in nerve fiber, and for cell are most important.Further, since sodium hyaluronate is that skin is true
Important moisturizing ingredient in cortex, and there is excellent glutinous capacitive and retractility, it is a kind of ideal filler, therefore often
For in beauty product and plastic operation.
Sodium hyaluronate is a glycosaminoglycan for not having element sulphur, and basic structure is by two dissacharide units D-Glucose aldehyde
The large-scale polysaccharide of acid and N-acetyl-glucosamine composition, the chemical formula of tool such as following formula (I):
Sodium hyaluronate can be prepared from natural materials, for example, by the living bodies such as cockscomb, umbilical cord, cartilage and skin partial extraction and
, it can also be produced by chemical synthesis or using microorganisms such as saccharomycete by culture or gene engineering method.Commercially available sodium hyaluronate
Preparation also can be used as the source of sodium hyaluronate in medical composition of the present invention, such as the preparation of the sodium water solution containing sodium hyaluronate, including
ARTZAndDeng, or comprising being crosslinked glass
The preparation of uric acid derivative aqueous solution, includingAnd
There is no particular restriction for the sodium hyaluronate of composition (a) suitable for medical composition of the present invention, but preferably has average
About 30,000 to about 10,000,000 dalton (Dalton) of molecular weight more preferably has about 500,000 to about 6,000,000 dongle of average molecular weight
?.In addition, sodium hyaluronate can be the free state of not forming salt, pharmaceutically permissible salt can also be formed.Sodium hyaluronate is in pharmaceutically
Permissible salt includes that salt is formed by with inorganic base, such as alkali such as the alkali metal ions such as sodium salt or sylvite salt, magnesium salts or calcium salt
Earthmetal cations salt and ammonium salt etc.；Or salt is formed by with organic base, such as with diethanol amine, cyclohexylamine or Amino acid etc.
It is formed by salt.
Vitamin (or vitamin) is micro-nutrient composition needed for organism, can not be produced by organism, it is necessary to
It is obtained by extrinsic pathways such as such as diet.Although vitamin as image of Buddha carbohydrate, protein and fat in organism without generating
Energy, but it has important adjustment effect to the metabolism of organism.The present inventor is the study found that such as rear attached embodiment institute
Show, compared to sodium hyaluronate or vitamin C being administered alone Ji ╱ or vitamin E, when merge using sodium hyaluronate and vitamin C Ji ╱ or
When vitamin E, the anti-inflammatory effect of enhancement can be unexpectedly provided, provides to be reached anti-so as to lower sodium hyaluronate dosage
Inflammation effect, the subsequent inflammatory response that may cause after slowing down, even sodium hyaluronate being avoided to enter human body.
In medical composition of the present invention, there is no particular restriction for the content proportion of composition (a) and (b).Composition (a) and (b)
Weight ratio be usually 1:0.02 to 1:1, preferably 1:0.05 to 1:0.5.
Medical composition of the invention can be used on animal doctor and human medical, and can be in any form, and with any conjunction
Suitable mode is applied.For example, but not limited to this, which can take orally, subcutaneously, vein or intra-articular injection
Equal dosing modes application.Depending on use form and purposes, a composition (c) medicine can be included in medical composition of the invention
Acceptable carrier.
It, can be in medical composition of the invention containing adversely for preparing and be suitable for the medicine type of oral administration medicine supplying
The medicine for influencing sodium hyaluronate and vitamin is subjected to carrier, such as: solvent, oil-based solvent, diluent, stabilization agent, absorption delay
Agent, disintegrating agent, emulsifier, antioxidant, binder, lubricant, hygroscopic agent etc..Using any suitable method, by the group
It closes object and the form suitable for oral administration medicine supplying is made, such as: pastille, capsule, granule, powder, liquid extract, solution, syrup
Agent, suspension agent, emulsion and tincture etc..
As for the medicine type for being suitable for subcutaneous, vein or intra-articular injection, then can contain in medical composition of the invention
Just like isotonic solution, water for injection, normal saline solution or salt buffer (such as phosphate buffer or citrate buffer)
Equal solvent, and one or more additive (such as hydrogen sulfates such as pH adjuster, isosmoticity agent or stabilization agent optionally
Sodium, double hydrosulphuric acid sodium, sodium dihydrogen phosphate, dibastic sodium phosphate or sodium chloride etc.), to be made such as venoclysis liquid, emulsion venoclysis
Liquid, dry powder injection, suspension injection or dry powder suspension injection etc..
Medical composition of the invention can be optionally in addition containing additives such as flavoring agent, toner, colorants, to improve
The suitable sense of mouth and visual experience when gained medicament is taken；Preservative agent appropriate, preservative, antibacterial agent, antifungal agent can also be added
Deng to improve the storage characteristics of gained medicament.
Optionally, in medical composition of the invention and one or more other active components can be contained, further to add
The effect of strong composition or the use flexibly and allotment degree for increasing pharmaceutical formulation.It for example, can be in of the invention medical group
It closes object and contains one or more following active constituents: steroids anti-inflammatory drug, on-steroidal anti-inflammatory drug, gucosamine
(glucosamine) and other active components etc., as long as other active components are to the benefit of sodium hyaluronate and vitamin without not
The influence of benefit.
It since medical composition of the invention can provide the anti-inflammatory effect of enhancement, therefore can be used for inhibiting inflammation, especially may be used
For inhibiting arthritis, including it is osteoarthritis (degenerative arthritis), rheumatoid arthritis, urarthritis, bacillary
Arthritis, ankylosing spondylarthritis and erythematosus lupus etc. are preferably used for inhibiting osteoarthritis and rheumatoid arthritis.In
In one embodiment, medical composition of the invention is made to the pattern of injection, for the treatment method for joint injection.
Significant therapeutic effect can not be reached for rheumatoid arthritis compared to existing sodium hyaluronate preparation, the present invention
Medical composition one be advantageous in that, can by inhibiting knee joint synovial bursa cell secreting tumor necrosis factor (TNF) (swollen
Tumor necrosis factor is the important virulence factor of rheumatoid arthritis), the effect of excellent inhibition rheumatoid arthritis is provided,
Therefore, it is used especially for the treatment of rheumatoid arthritis.In addition, medical composition of the invention can also be by inhibiting inflammation phase
The performance of the factor (such as IL-1, IL-6, IL-8) is closed, the secretion of anti-inflammatory cytohormone (IL-10) is promoted and promotes structure phase
The performance of protein (such as MMP-3 shows the marker of synovial membrane inflammation) is closed to achieve the effect that inhibit inflammation.Degenerative joint
Inflammation is considered as a kind of atypical inflammatory disease in recent years, by inhibiting first type Jie Bai Su, can also slow down degenerative joint
Scorching progress, and then achieve the purpose that treat degenerative arthritis.
It is outside one's consideration in addition, the present composition removes the arthralgia that can effectively mitigate patient and improves joint motion, it can also be pre-
Prevent arthritic generation.Also, it is different from the combination of known sodium hyaluronate and aminomethyl folic acid, the complicated hand of victory peptide must be passed through
Section to form conjugate can provide the anti-inflammatory effect to be reached, in medical composition of the present invention, sodium hyaluronate and opposite
A small amount of vitamin C and/or vitamin E only needs to be simply mixed, therefore has simple process, suitable for the excellent of scale of mass production
Point.
Since medical composition of the present invention can improve the defect of existing sodium hyaluronate preparation, therefore apply also for sodium hyaluronate
Any known purposes, and it is not limited to Antiarthritic.For example, the present composition can be applied to outside beauty product or shaping
Section, such as can be made an addition in skin care product or the sodium hyaluronate injection of face.
The present invention also provides a kind of methods for inhibiting inflammation, and it includes so that a patient is applied an a effective amount of doctor of the invention
Drug composition.It is the pattern with injection in one embodiment of the invention, medical composition of the present invention is injected to patient's
In joint, to reach the effect for the treatment of of arthritis.
The present invention also provides a kind of for inhibiting the set group of inflammation comprising: the first of a sodium hyaluronate containing effective dose
The second part of part and the vitamin containing effective dose.Wherein, the type of sodium hyaluronate and vitamin, property in the set group
Matter, source and usage ratio etc. are as described above.
In set group of the present invention, the form of the first part and second part has no specifically limited.For example, in for closing
In section in the set group of injection dispensing, the first part and second part can be in solution form and be respectively placed in different sterilization packagings
In (such as plastic bottle or vial such as ampulla), and each packaging may include multiple dosage but preferably unitary dosages agent
The first part of amount or second part.In this, set group of the present invention may also include an an injection needle tube (such as deserted injection needle
Cylinder), or optionally further include an operation instructions.It, can be according to the instruction of the operation instructions when carrying out intra-articular injection dispensing
(information such as ratio mixed including such as mode of operation of set group and solution), two parts are fitted into the injection needle tube to apply medicine
Agent.
On the other hand, in the set group for oral administration medicine supplying, the first part and second part can respectively independently in pastille or
The form of capsule, and the two parts can be respectively placed in Different Package according to unitary dosages dosage or multiple dosage.In this, originally
Invention set group can optionally include an operation instructions, when Yu Jinhang oral administration medicine supplying, can according to the operation instructions instruction (including
Time-histories of such as offeing medicine and recommended doses information), merge the two parts and is taken.
Now with following specific examples further to illustrate the present invention.Wherein these embodiments are merely provided as
Protection scope that is bright, and being not intended to limit the invention.
[preparation embodiment]
A, the preparation of sodium hyaluronate joint injection liquid
In in a prepackage type sodium hyaluronate injection needle device, (average molecular weight is for sodium hyaluronate that addition concentration is 0.5 to 2 milligram
60 to 800,000 dalton) 1 milliliter and 0.01 to 1 milligram of isotonic solution of vitamin C or vitamin E, then total amount of adding is 5 to 20
Sodium chloride, sodium bisulfate and the double hydrosulphuric acid sodium of milligram finally add 2 milliliters of waters for injection as excipients, glass urine is made
Sour joint injection liquid is placed under 4 DEG C of light protected environment and stores.
B, the preparation of fibroblast sample synovial cell
In arthroscope debridement surgical, collects and suffered from degenerative arthritis (osteoarthritis, OA) from five
Sufferer fibroblast sample synovial cell (fibroblast-like synoviocytes, FLS) and cultivated.It is first
First, the synovium of joint (synovium) of sufferer is cut into fractionlet, then it is made to be suspended in a DMEM (Dulbecco modified
Eagle ' s medium) in culture medium, and cultivated 3 days in 37 DEG C, the environment of 5% carbon dioxide.The culture medium contains 1.5
G/liter sodium bicarbonate (S6297, Sigma-Aldrich, St Louis, the Missouri State, the U.S.), 1% penicillin-chain
Mycin-neomycin (P4083, Sigma-Aldrich) and 10% fetal calf serum (04-001-1A, Biological
Industries, Grand Island, New York, the U.S.).
Non-adhering cells are washed away with phosphate buffer (PBS), and replace culture medium, are further cultured for the thin of the attachment left
Born of the same parents last 2 weeks.Repeat above procedure 3 to 6 times, the cell under retaining is fibroblast sample synovial cell, be used for
Lower test.
The test cell line of [embodiment 1] sodium hyaluronate joint injection liquid
The processing of (1-1) cell
Fibroblast sample synovial cell obtained by culture [preparation embodiment B] goes through in the culture medium without containing serum
When 24 hours, then respectively at being cultivated 24 hours in following four kinds of culture mediums: (1) the only DMEM culture containing 10% fetal calf serum
Base, this is as control group；2) (average molecular weight is 60 to 800,000 for the DMEM culture medium containing 10% fetal calf serum and sodium hyaluronate
Er Dun)；3) the DMEM culture medium containing 10% fetal calf serum and vitamin C or vitamin E；And 4) contain 10% fetal calf serum
DMEM culture medium, 100 mcg/mls sodium hyaluronate (average molecular weight be 60 to 800,000 dalton) and 50 mcg/mls
Vitamin C or 25 micro- Moore concentration (that is, 10.76 mcg/mls) vitamin E (this that is, sodium hyaluronate with it is ascorbic heavy
Amount is than being about sodium hyaluronate: vitamin C=1:0.5；The weight ratio of sodium hyaluronate and vitamin E is about sodium hyaluronate: vitamin E=1:
0.1)。
Then, above-mentioned four groups of cells are collected respectively and are centrifuged, and supernatant are regathered, to carry out follow-up test.
(1-2) measures quantifying for protein relevant to arthritis by ELISA
Inflammatory response is a complicated process, wherein comprising generate free radicals molecule (such as: nitric oxide and peroxidating
Hydrogen etc.) and cytohormone (cytokine) (such as: prostaglandin 2 (PGE2), tumor necrosis factor-alpha (TNF-α), interferon-
γ (IFN-γ), Jie Bai Su -2 (IL-2, Interleukin-2) and Jie Bai Su -1 β (IL-1 β)) etc..Therefore, by measurement with
The relevant free radical molecule of inflammatory response or cytohormone, it can be learnt that the degree of inflammatory response.
In this experiment, set group (sandwich binding protein assay is analyzed using interlayer conjugated protein
Kit or sandwich ELISA kits is purchased from eBioscience company), and the service manual according to manufacturer, utilize standard song
Line is to measure the concentration of two cytohormones relevant to degenerative arthritis in supernatant collected by 1-1, to observe its performance
Degree, judges the inflammation situation of cell whereby, and two kinds of cytohormones are respectively that (standard items are purchased from 88- to tumor necrosis factor-alpha
7340, eBioscience) and Jie Bai Su -1 β (88-7010, standard items be purchased from eBioscience company).Each sample standard deviation carries out
It tests, and is measured using ferment immuning adsorpting analysis (ELISA) reader (Sunrise Remote, TECAN) twice.
As a result shown in table 1, table 2 and Fig. 1 to Fig. 4.
Table 1
Table 2
(1-2) test result
Show that the joint cell from degenerative arthritis patient can secrete a large amount of hair by table 1, table 2 and Fig. 1 to Fig. 4
Scorching medium T NF- α and IL-1 β illustrates that its inflammation situation is serious, and combines sodium hyaluronate and vitamin C and/or vitamin E, then may be used
Effectively inhibit the performance of the inflammation medium.Wherein, as shown in Figures 1 and 3, present invention combination sodium hyaluronate and vitamin C and/or dimension
TNF-α (the important virulence factor of rheumatoid arthritis) performance can be effectively suppressed in raw element E, therefore can provide excellent inhibition and close
The effect of saving inflammation, especially rheumatoid arthritis.In addition, as shown in Figures 2 and 4, present invention combination sodium hyaluronate and vitamin C
And/or vitamin E, it can also be by inhibiting IL-1 β performance to reach the effect of inhibition inflammation, especially inhibition degenerative arthritis
Fruit.The above test data, also showing medical composition of the present invention can provide not expected enhancement anti-inflammatory effect.
[embodiment 2] sodium hyaluronate or vitamin C is used alone Ji ╱ or vitamin E and and with sodium hyaluronate and vitamin C
And/or vitamin E, in the anti-inflammatory the effect comparison of
The processing of (1-3) cell
(1-4) (2-1-1) sodium hyaluronate and with vitamin C group
Fibroblast sample synovial cell obtained by culture [preparation embodiment B] reaches in the culture medium without containing serum
After 24 hours, be incubated in the DMEM culture medium containing 10% fetal calf serum, and be divided into following eight groups: 1) holding cell is
Do not receive the control group for the state for stimulating and handling；2) individually with 100 mcg/ml sodium hyaluronates, (abbreviation HA, average molecular weight are
60 to 800,000 dalton) cell is handled up to 24 hours；3) cell is individually handled with 50 mcg/ml vitamin Cs (abbreviation Vit C)
Up to 24 hours；4) with the sodium hyaluronate of 100 mcg/mls and 50 mcg/ml vitamin Cs processing cell up to 24 hours；5) it is received with 1
Grams per milliliter IL-1 β (4128-10, BioVision, the U.S.) stimulates cell up to 24 hours, and (wherein, IL-1 β can stimulate cell for one
Jie's white matter of inflammation)；6) with 1 nanograms/milliliter IL-1 β stimulation cell 24 hours after, individually with 100 mcg/ml sodium hyaluronates at
Cell is managed up to 24 hours；7) with 1 nanograms/milliliter IL-1 β stimulation cell 24 hours after, individually with 50 mcg/ml vitamin Cs at
Cell is managed up to 24 hours；8) after with 1 nanograms/milliliter IL-1 β stimulation cell 24 hours, with the sodium hyaluronate of 100 mcg/mls and 50
Mcg/ml vitamin C handled cell up to 24 hours.
Then, above-mentioned eight groups of cells are collected respectively and are centrifuged, and supernatant are regathered, to carry out follow-up test.
(2-1-2) sodium hyaluronate and with vitamin E group
Fibroblast sample synovial cell obtained by culture [preparation embodiment B] reaches in the culture medium without containing serum
After 24 hours, be incubated in the DMEM culture medium containing 10% fetal calf serum, and be divided into following four groups: 1) holding cell is
Do not receive the control group for the state for stimulating and handling；2) individually with 100 mcg/ml sodium hyaluronates, (abbreviation HA, average molecular weight are
60 to 800,000 dalton) cell is handled up to 24 hours；3) it is individually handled with 25 micro-molar concentration vitamin Es (abbreviation Vit E) thin
Born of the same parents were up to 24 hours；4) with the sodium hyaluronate of 100 mcg/mls and 25 micro-molar concentration vitamin Es processing cell up to 24 hours.
Then, above-mentioned four groups of cells are collected respectively and are centrifuged, and supernatant are regathered, to carry out follow-up test.
(2-2) measures quantifying for protein relevant to arthritis by ELISA
It is analyzed according to the test method of 1-2, four inflammation are relevant in collected supernatant in measurement 2-1 each group
The concentration of the factor judges that the inflammation situation of cell, the factor are respectively Jie Bai Su -1 β (IL-1 to observe its performance degree whereby
β, standard items are purchased from 88-7010, eBioscience), Jie Bai Su -8 (IL-8, standard items be purchased from DY208, R&D systems),
Transin-1 (MMP-3, standard items be purchased from DY513, R&D systems) and tumor necrosis factor-alpha (TNF-α,
Standard items are purchased from 88-7340, eBioscience).Each sample standard deviation is tested twice, and uses a ferment immuning adsorpting analysis
(ELISA) reader (Sunrise Remote, TECAN) is measured.Double sample t inspection (two- is carried out to the data obtained
Sample t-test), as a result it is shown in table 3-1 to 3-4, table 4-1 to 4-4, Fig. 5-1 to 5-4 and Fig. 6-1 to 6-4.
Wherein, table 3-1 to 3-4 and table 4-1 to 4-4 is classified as the average value and inflammation correlation factor of each test data
For mean concentration compared to the decline percentage of control group, Fig. 5-1 to 5-4 and Fig. 6-1 to 6-4, which is then shown with histogram, tests knot
Fruit.
Table 3-1
Table 3-2
Table 3-3
Table 3-4
Table 4-1
Table 4-2
Table 4-3
Table 4-4
(2-2) test result
As shown in table 3-1 to 3-4, table 4-1 to 4-4, Fig. 5-1 to 5-4, Fig. 6-1 to 6-4, suffer from from degenerative arthritis
The joint cell of person can secrete a large amount of inflammation medium IL-1, IL-8, MMP-3 and TNF-α, illustrate that its inflammation situation is serious.And
If the present invention combines sodium hyaluronate and vitamin C and/or vitamin E, then IL-1, IL-8, MMP-3 and TNF-α can be effectively suppressed
The effect of showing, therefore can provide excellent inhibition arthritis, especially rheumatoid arthritis.In addition, compared to exclusive use
Sodium hyaluronate or vitamin C and/or vitamin E handle cell, and present invention combination sodium hyaluronate can be mentioned with vitamin C and/or vitamin E
The effect of being showed for more excellent inhibition IL-1, IL-8, MMP-3 and TNF-α.
Wherein, such as shown in table 3-1 and Fig. 5-1, when sodium hyaluronate processing cell is used alone, the IL-1's of cell is averaged
Concentration be 6.28 ± 0.72 pico- grams per milliliters, compared to control group concentration decline percentage be 8.18% (calculation:
6.84-6.28/6.84 X 100%=8.18%)；When vitamin C processing cell is used alone, the IL-1's of cell is average dense
Degree then be 6.34 ± 0.73 pico- grams per milliliters, compared to control group concentration decline percentage be 7.30% (calculation:
6.84-6.34/6.84 X 100%=7.30%)；And when sodium hyaluronate is with vitamin C processing cell, the IL-1's of cell
Mean concentration is then reduced to 4.11 ± 1.09 pico- grams per milliliters, and the concentration decline percentage compared to control group is then up to 39.91%
(6.84-4.11/6.84 X 100%=39.91%), this numerical value, which is apparently higher than, is used alone sodium hyaluronate or vitamin C processing carefully
Born of the same parents, the summation result of even higher than two experimental groups.Aforementioned experimental results are shown, compared to exclusive use sodium hyaluronate or vitamin
C, the present invention and with sodium hyaluronate and vitamin C processing cell when, it is possible to provide have not expected multiplication effect
The inhibition inflammation effect of (synergistic effect, the i.e. effect of 1+1 > 2).
Such as shown in table 4-1 and Fig. 6-1, when sodium hyaluronate processing cell is used alone, the mean concentration of the IL-1 of cell
For 7.10 ± 1.02 pico- grams per milliliters, the concentration decline percentage compared to control group is 12.01% (calculation: 8.07-
7.10/8.07 X 100%=12.01%)；When vitamin E processing cell is used alone, the mean concentration of the IL-1 of cell is then
For 6.12 ± 0.65 pico- grams per milliliters, the concentration decline percentage compared to control group is 24.16% (calculation: 8.07-
6.12/8.07 X 100%=24.16%)；And when sodium hyaluronate is with vitamin E processing cell, the IL-1's of cell is averaged
Concentration is then reduced to 1.91 ± 0.51 pico- grams per milliliters, and the concentration decline percentage compared to control group is then up to 76.33%
(8.07-1.91/8.07 X 100%=76.33%), this numerical value, which is apparently higher than, is used alone sodium hyaluronate or vitamin E processing carefully
Born of the same parents, the summation result of even higher than two experimental groups.Aforementioned experimental results are shown, compared to exclusive use sodium hyaluronate or vitamin
E, the present invention and with sodium hyaluronate and vitamin E processing cell when, it is possible to provide with not expected multiplication effect inhibition inflammation
Effect.
[embodiment 3] and with sodium hyaluronate and relatively great amount of vitamin C and/or vitamin E and and with sodium hyaluronate and phase
To a small amount of vitamin C and/or vitamin E, in the anti-inflammatory the effect comparison of
The processing of (3-1) cell
Fibroblast sample synovial cell obtained by culture [preparation embodiment B] reaches in the culture medium without containing serum
After 24 hours, then respectively at being cultivated in the DMEM culture medium containing 10% fetal calf serum with following condition:
1st group: micro- with the sodium hyaluronate (abbreviation HA, average molecular weight are 60 to 800,000 dalton) of 100 mcg/mls and 50
Vitamin C (abbreviation Vit C is purchased from Sigma, article No. A8960, the U.S.) processing of grams per milliliter, cultivates 24 hours (that is, sodium hyaluronate
It is sodium hyaluronate: vitamin C=1:0.5 with ascorbic weight ratio)；Or
2nd group: with the sodium hyaluronate (average molecular weight is 60 to 800,000 dalton) and 200 micrograms/milli of 100 mcg/mls
Vitamin C (being purchased from Sigma, article No. A8960, the U.S.) processing risen, culture 24 hours (that is, sodium hyaluronate and ascorbic weight
Than for sodium hyaluronate: vitamin C=0.5:1).
Then, above-mentioned two groups of cells are collected respectively and are centrifuged, and supernatant are regathered, to carry out follow-up test.
(3-2) measures the performance amount of protein relevant to degenerative arthritis by ELISA
It is analyzed according to the test method of 1-2, measures in 3-1 each group in collected supernatant five and closed with degenerative
The concentration of the scorching relevant factor of section judges that the inflammation situation of cell, the factor are respectively as follows: (I) to observe its performance degree whereby
Inflammation relevant cell hormone: IL-6 (88-7066, eBioscience, San Diego, Canada), IL-8 (DY208, R&D
Systems, the U.S.) and TNF-α (88-7340, eBioscience, the U.S.)；(II) anti-inflammatory cytohormone: IL-10 (88-
7106, eBioscience, the U.S.)；(III) structure related protein: MMP-3 (that is, marker of display synovial membrane inflammation)
(DY513；R&D System, the U.S.).Each sample standard deviation is tested twice, and uses a ferment immuning adsorpting analysis (ELISA)
Reader (Sunrise Remote, TECAN) is measured.Double sample t inspection (two-sample t- is carried out to the data obtained
Test), table 5 and Fig. 7-1 are as a result shown in 7-5.Wherein, table 5 is shown as the statistical result of average value ± standard deviation.
Table 5
(3-2) test result
As table 5 with and Fig. 7-1 to 7-5, compared to the 1st group cell (being handled through HA:Vit C=1:0.5), the 2nd group of cell
(handling through HA:Vit C=0.5:1) secretes more inflammation relevant cell hormone (IL-6, IL-8), less anti-inflammatory cell swashs
Plain (IL-10) and more structure related protein (MMP-3, that is, the marker of display synovial membrane inflammation).Aforementioned result is aobvious
Show, compared to and with sodium hyaluronate and relatively small amount vitamin C (for example, HA:Vit C=1:0.5), and with sodium hyaluronate and opposite
The anti-inflammatory effect of a large amount of vitamin C (for example, HA:Vit C=0.5:1) is obviously poor, shows and with sodium hyaluronate and relatively
A large amount of vitamin C can not provide the anti-arthritic efficacy with synergistic effect (synergistic effect).This test knot
Fruit, it was demonstrated that the sodium hyaluronate that the present invention is defined has critical with the numerical definiteness range of vitamin C and/or vitamin E amount ratio
Meaning, the present invention can provide really can not expected beneficial functional easily.
Above-described embodiment is only to illustrate the principle of the present invention and effect, and is not intended to limit the present invention.This field
Technical staff can modify to above-described embodiment and become without prejudice to technical principle and spirit of the invention
Change.Therefore, protection scope of the present invention is answered subject to claims define.
Claims (10)
1. a kind of for inhibiting the medical composition of inflammation, which is characterized in that it includes:
(a) sodium hyaluronate；
(b) vitamin；And
(c) the acceptable carrier of medicine；
Wherein, composition (b) is at least one of vitamin C and vitamin E；And the weight ratio of composition (a) and (b) are 1:0.02
To 1:1.
2. as described in claim 1 for inhibiting the medical composition of inflammation, which is characterized in that the mean molecule of composition (a)
Amount is about 30,000 to about 10,000,000 dalton (Dalton).
3. as described in claim 1 for inhibiting the medical composition of inflammation, which is characterized in that the mean molecule of composition (a)
Amount is about 500,000 to about 6,000,000 dalton.
4. as described in claim 1 for inhibiting the medical composition of inflammation, which is characterized in that the weight of composition (a) and (b)
Than for 1:0.05 to 1:0.5.
5. according to any one of claims 1 to 4 for inhibiting the medical composition of inflammation, which is characterized in that the use
In inhibit the medical composition of inflammation be in for oral, subcutaneous injection, intravenous injection or intra-articular injection dispensing in the form of.
6. according to any one of claims 1 to 4 for inhibiting the medical composition of inflammation, which is characterized in that the use
Antiarthritic is used in the medical composition of inhibition inflammation.
7. as claimed in claim 6 for inhibiting the medical composition of inflammation, which is characterized in that described for inhibiting inflammation
Medical composition is used for anti-osteoarthritis, resisting rheumatoid arthritis or antigout arthritis.
8. as claimed in claim 7 for inhibiting the medical composition of inflammation, which is characterized in that described for inhibiting inflammation
Medical composition is used for resisting rheumatoid arthritis.
9. a kind of for inhibiting the set group of inflammation, characterized in that it comprises:
One first part, the sodium hyaluronate comprising effective dose；And
One second part, the vitamin comprising effective dose；
Wherein, which is at least one of vitamin C and vitamin E；And the weight ratio of the sodium hyaluronate and the vitamin
For 1:0.02 to 1:1.
10. as claimed in claim 9 for inhibiting the set group of inflammation, which is characterized in that the first part and second part are in
For the solution form of injection dispensing, and the set group further includes an injection needle tube.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101983722A (en) * | 2010-10-20 | 2011-03-09 | 北京康比特体育科技股份有限公司 | Composition for preventing and treating joint soft tissue injury and complications thereof |
| CN102988406A (en) * | 2011-09-16 | 2013-03-27 | 中国医药大学 | Pharmaceutical composition and kit for inhibiting inflammation |
| CN106794140A (en) * | 2014-05-30 | 2017-05-31 | 娜塔莉亚·米克拉维娜·德卡奇 | Pharmaceutical composition based on active material stablizing solution |
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2018
- 2018-01-16 CN CN201810041014.6A patent/CN110038027A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101983722A (en) * | 2010-10-20 | 2011-03-09 | 北京康比特体育科技股份有限公司 | Composition for preventing and treating joint soft tissue injury and complications thereof |
| CN102988406A (en) * | 2011-09-16 | 2013-03-27 | 中国医药大学 | Pharmaceutical composition and kit for inhibiting inflammation |
| CN106794140A (en) * | 2014-05-30 | 2017-05-31 | 娜塔莉亚·米克拉维娜·德卡奇 | Pharmaceutical composition based on active material stablizing solution |
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