CN101247834A - A carrier comprising one or more di and/or mono-(electron transfer agent) phosphate derivatives or complexes thereof - Google Patents
A carrier comprising one or more di and/or mono-(electron transfer agent) phosphate derivatives or complexes thereof Download PDFInfo
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- CN101247834A CN101247834A CN200680021329.XA CN200680021329A CN101247834A CN 101247834 A CN101247834 A CN 101247834A CN 200680021329 A CN200680021329 A CN 200680021329A CN 101247834 A CN101247834 A CN 101247834A
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Abstract
The invention relates to a carrier for administering biologically active compounds comprising one or more C1-C4 alcohols, polyols and polymers thereof, water and one or more di and/or mono-(electron transfer agent) phosphate derivatives or complexes thereof. The carrier may be used in administering biologically active compounds, in particular pharmaceuticals including cosmetic agents.
Description
Technical field
The present invention relates to be used to use the carrier of biologically active cpds and contain biologically active cpds and the preparation of described carrier.Described carrier help to improve described biological compound (medicine that particularly comprises enamel) effectiveness, transport and send.
Background technology
In this manual, when quoting or when discussion paper, behavior or article, this is quoted or discuss and is not to recognize that: the knowledge of described file, behavior or article or its be any, and to be combined in preference day be that the public can be known, the part of known, common practise; Perhaps known effort with the described any problem of this description of solution is associated.
Main purpose during medicine is sent is the suitable biological effect of site of action acquisition in hope.The selection of preparation can be crucial for pharmaceutical efficacy, because if medicine does not have suitable plysiochemical character so that it discharges from preparation at target site, then its biological activity is not good enough.
Intestinal delivery comprises that medicine is absorbed therein, and is distributed to target site by blood flow by gastrointestinal (GI) road drug administration.For example, the medicine of oral delivery is via intestinal absorption.
It also is important that the chemical environment in GI road is sent for external drug.Medicament forms must be stable under the different pH of the various piece in GI road.If medicine forms non-absorbent complex or chemical degradation or enzymatic degradation, will reduce absorption.Medicine also must be the solution in GI liquid, to be absorbed.The precipitation of medicine comprises that medicine forms solid particle and therefore breaks away from solution.The solid particle that is adsorbed to official jargon comprises the solid that absorbs the drug; That is, from solution, remove described medicine.Drug precipitation and absorption have all reduced the absorption of medicine.In many cases, degraded and complexation can prevent or minimizing at least by chemistry or formulation method, so that there is not the restriction to drug absorption in it.
And if medicine absorbs by intestinal wall or coat of the stomach, then it must pass through liver.The effect of liver is to remove xenobiontics from body.Therefore, significantly the medicine of ratio (for example, 40-50%) may and be drained by metabolism before arriving blood flow.By medicine is absorbed via mouth internal layer (oral cavity (bucchal)/Sublingual) or rectum internal layer (suppository), may reduce the influence of liver, but these approach are not always suitable to the enteral administration.
The effort that improves the bioavailability of medicament of enteral administration comprises the excipient that forms prodrug (for example, morphine sulfate) or use enhancing to absorb.
Topical comprises that with drug administration to the body film, medicine is absorbed and distributes in the body film.For example, the medicine of percutaneous dosing is via skin absorbs.
Skin is the organ of body maximum, and it does to avoid outside chemistry, physics and pathology harm in order to the protection internal.Normal skin is divided into three layers: epidermis, corium and subcutaneous tissue.That the cornified skin (horny layer) of epidermis has is tough, elasticity, high resistance and exsiccant character, and it stops penetrating of microorganism and breeds.Horny layer also is the major obstacle that transdermal drug absorbs.Have the sebum layer of protection skin, it is considered to all obstacles based on the pharmaceutical preparation of water.
When passing skin, the drug molecule of diffusion has three potential approach that enter the deep skin layer: iuntercellular approach, transcellular pathway and through adnexa approach (transappendageal route).Though electrolyte and macromole are significant via the bypass of adnexa diffusion, the relatively little area that can be used for transporting (skin surface 0.1%) means that this approach has insignificant contribution to the drug flow of steady statue.Generally believe that the main path that is used for through most molecules is the iuntercellular approach, therefore, many enhancement techniques are devoted to destroy cuticular strong " fragment of brick and mortar " structure.Point to two kinds of possible mechanism about the theory of transporting pathway at present: (i) passive transcellular and (ii) cell endepidermis transhipment.
Medicine is applied topically to skin in many ways, and described mode comprises: ointment, patch, solution, subcutaneous storage agent, poultice, plaster and transdermal delivery device.
May be to the concern of transdermal drug delivery in continuous increase, but some basic restrictions have restricted the broader applications of this technology.The major limitation of using transdermal administration is the transport velocity of medicine via skin.
Be not that every kind of medicine can both be with sufficiently high speed percutaneous dosing, treatment has the blood drug level for the treatment of benefit to systemic drug to obtain.For example, the medicine with similar molecular wt and size can be striden skin absorbs by different rates.For example, fentanyl is with 2mg/cm
2/ hr sees through skin, and ephedrine is with 200mg/cm
2/ hr.Therefore, although have the advantage of route of administration, the needed large scale transdermal delivery system of fentanyl is both impracticable also uneconomical.
Developed the percutaneous absorption that skin reinforcing agent and various preparation technique improve medicine.The skin reinforcing agent can comprise that as chemical compounds such as capric acid, oleic acid, azone, decyl methyl sulfoxide and hydroxycinnamic acid salt it is done in order to change particularly cuticular structure, to improve the medicine permeability by the dissolving lipid matrix usually.For example, when horny layer during by degrease, the epidermis of Progesterone absorbs increases by 143%.When horny layer was removed fully, potentiation was increased to 843%.Because this aggressivity changes, widespread reports to follow the reusable problem of this system be obvious, comprise scratching where it itches and carbonization of contact dermatitis, skin reddening, the mobile patch of needs, perhaps all body drug application are to prevent local excitation.It is said that reddening is being removed paster in a few hours hour.But produced long-term risk and safe concern to using the type transdermal delivery system, main because the medicine permeability that improves is a cost with the most important protective layer of infringement skin.
Existence is to the demand of the preparation of further raising bioactive compound bioavailability.
Summary of the invention
Summary of the invention
Have been found that if bioactive compound with the carrier administration, then its effectiveness, transport and send and can improve, described carrier comprises: one or more C
1-C
4Alcohol, polyhydric alcohol and polymer thereof; Water; With one or more two-and/or list-(electron transfer agent) phosphate derivative or its complex.
According to a first aspect of the invention, provide the carrier that is used to use bioactive compound, this carrier comprises: one or more C
1-C
4Alcohol, polyhydric alcohol and polymer thereof; Water; With one or more two-and/or list-(electron transfer agent) phosphate derivative or its complex.
The present invention also provides one or more C
1-C
4Alcohol, polyhydric alcohol and polymer thereof, water and one or more two-and/or list-(electron transfer agent) phosphate derivative or its complex be used for using the purposes of the carrier of bioactive compound in production.
The method for preparing above-mentioned carrier also is provided, and it comprises the steps:
(a) with one or more two-and/or list-(electron transfer agent) phosphate derivative or its complex and one or more C
1-4Alcohol, polyhydric alcohol or its polymer combination; And
(b) water is added in the combination of step (a).
Should be understood that described carrier can prepare from alcohol, the sub-transfer agent phosphate derivative of power and water or its complex, or the product of alcohol, the sub-transfer agent phosphate derivative of power and water or its complex.In these cases, alcohol, the sub-transfer agent phosphate derivative of power and water or its complex can interact and exist with modified form.
Preferably, C
1-4Alcohol is ethanol.
Described carrier preferably contains the combination of one or more two-(electron transfer agent) phosphate derivatives or one or more two-(electron transfer agent) phosphate derivatives and one or more list-(electron transfer agent) phosphate derivatives.
Should be understood that term " two-and/or list-(electron transfer agent) phosphate derivative " refers to the phosphate ester of electron transfer agent, wherein phosphate ester can be by electron transfer agent two-or the orthophosphate or the pyrophosphate of list-replacement.
In one embodiment, two-(electron transfer agent) phosphate derivatives are selected from two-tocopherol phosphate derivative, two-tocopherol, two-phosphate derivative, two-tocotrienol phosphate derivative and composition thereof.Preferably, two-(electron transfer agent) phosphate derivatives are two-tocopherol phosphate esters.
List-(electron transfer agent) phosphate derivative is preferably selected from list-tocopherol phosphate derivative, list-tocopherol two-phosphate derivative, list-tocotrienol phosphate ester (tocotrienylphosphate) and composition thereof.
In a preferred embodiment, use at least a preparation preparation of two-tocopherol phosphate ester, two-tocopherol, two-phosphate ester and two-tocotrienol phosphate ester.
In another preferred embodiment, use following combined preparation preparation: at least a and two-tocopherol phosphate ester, two-tocopherol bisphosphate and two-tocotrienol phosphate ester of list-tocopherol phosphate ester, list-tocopherol two-phosphate ester and list-tocotrienol phosphate ester at least a.
When preparation contained the combination of list-tocopherol phosphate ester and two-tocopherol phosphate ester, these chemical compounds can its one or more α, β, γ and δ form exist preferred α and γ form.
The ratio of list-tocopherol phosphate ester and two-tocopherol phosphate ester is preferably 4: 1 to 1: 4, more preferably 2: 1.
The present invention further provides the preparation that comprises bioactive compound and carrier, described carrier comprises: one or more C
1-C
4Alcohol, polyhydric alcohol and polymer thereof; Water; With one or more two-and or list-(electron transfer agent) phosphate derivative or its complex.
According to the present invention, the method that is used to use bioactive compound further is provided, it comprises that described carrier comprises with bioactive compound and carrier-bound step: one or more C
1-C
4Alcohol, polyhydric alcohol and polymer thereof; Water; With one or more two-and or list-(electron transfer agent) phosphate derivative or its complex.
Described carrier can be the vesicle form.Bioactive compound can be wrapped up by vesicle to small part.Though do not think bound by theoryly, think the formation of vesicle with controlled malleability, can make preparation by the iuntercellular approach and will be delivered to target cell in the bioactive compound cell or enter systemic circulation.Two-and or list-(electron transfer agent) phosphate derivative help to resist the inflammation that causes by the preparation administration.
Describe in detail
Carrier of the present invention contains: one or more C
1-C
4Alcohol, polyhydric alcohol and polymer thereof; Water; With one or more two-and/or list-(electron transfer agent) phosphate derivative or its complex.Preferably, the water content scope is 50% to 99%, more preferably 60% to 95%, most preferably 70% to 90%.
Alcohol
Term " C
1-C
4Alcohol " refer to have the alcohol of 1 to 4 carbon atom, as C
1-4Alkanol (for example methanol, ethanol, propanol, isopropyl alcohol or butanols).Polyhydric alcohol and C
1-4Alkoxide polymer comprises dihydroxylic alcohols, as propylene glycol or Polyethylene Glycol (for example PEG400).Can also use the combination of alcohol.Ethanol is preferred.
C
1-C
4The content range of alcohol is preferably 0.5% to 50%, and more preferably 5% to 40%, most preferably 10% to 30%.
The electron transfer agent phosphate derivative
Term " electron transfer agent " refers to and can be accepted electronics to generate metastable molecule free radical or to accept two electronics so that this material sedimentary material in the reversible redox system by the material of phosphorylation and (with the non-phosphorylating form).Can be comprised by the example of the electron transfer agent of phosphorylation: hydroxychroman, as α, β, γ and the δ tocol of enantiomer and racemic form; Hydroquinone for electron transfer agent K1 and ubiquinone reduction form; Hydroxy kind carotene is as retinol; Calciferol and ascorbic acid.Preferably, electron transfer agent is selected from tocol, retinol, is hydroquinone of electron transfer agent K1 reduction form and composition thereof.
More preferably, electron transfer agent is a tocol, as tocopherol or tocotrienol.Tocol comprise have following formula (I) 6: hydroxyl 2: all isomers of methyl chroman derivatives comprise α-5:7:8 trimethyl, β-5:8 dimethyl, γ-7:8 dimethyl and δ 8 methyl-derivatives.
Wherein
R
1, R
2And R
3Independently be selected from hydrogen and C
1-4Alkyl, preferable methyl.
In tocopherol, R
4Be 4:8:12 trimethyl tridecane, and 2,4 and 8 (referring to *) have R or S activity or racemic stereoisomer.In tocotrienol, R
4Be 4:8:12 trimethyl tridecyl-3:7:11 triolefin, and 2 can be to have R or S activity or racemic stereoisomer.Most preferably, electron transfer agent is alpha-tocopherol or tocotrienol.
Term " phosphoric acid derivatives " refer to the sour form, phosphate (comprise slaine, as alkali metal salt or alkali salt, for example sodium salt, magnesium salt, potassium salt and calcium salt) of phosphorylation electron transfer agent and wherein the phosphoric acid proton by other substituent groups (as C
1-C
4Alkyl or phosphatidyl) any other derivant of replacing.
Under the certain situation, may use phosphate derivative, as phospholipid.Phosphatidyl derivant is the aminoalkyl derivant of phosphate ester.These derivants can be R from structure
5R
6N (CH
2)
nThe amine preparation of OH, wherein n is 1 to 6 integer, and R
5And R
6Independently be selected from H and C
1-4Alkyl.Phosphatidyl derivant is prepared as follows: with the hydroxyl proton of phosphoric acid entity replacement electron transfer agent, and phosphoric acid entity and amine (as ethanolamine or N, N ' dimethylethanolamine) reaction then.A kind of method for preparing phosphatidyl derivant comprises: basic solvent (as pyridine or triethylamine) prepares intermediate with phosphorous oxychloride, the hydroxyl reaction of this intermediate and amine is to generate the corresponding phospholipids acyl derivative, as P-gallbladder acyl-P (P cholylP) biphosphate tocopherol then.
Term " C
1-4Alkyl " refer to have straight chain, straight chain or the cyclic hydrocarbon group of 1 to 4 carbon atom.Example comprises: methyl, ethyl, propyl group, isopropyl, butyl, isobutyl group, sec-butyl, the tert-butyl group, cyclopropyl and cyclobutyl.
Particularly preferred electron transfer agent phosphate derivative is two-tocopherol phosphate derivative, two-tocopherol, two-phosphate derivative, two-tocotrienol phosphate derivative, list-tocopherol phosphate derivative, list-tocopherol two-phosphate derivative and list-tocotrienol phosphate derivative, the combination of optimum menu-tocopherol phosphate derivative and two-tocopherol phosphate derivative.
The stability that has been found that carrier increases with the increase of list-electron transfer agent (list-alpha-tocopherol phosphate ester) concentration.When having the combination of list-alpha-tocopherol phosphate ester and two-tocopherol phosphate ester, its preferred proportion is 4: 1 to 1: 4, more preferably 2: 1.
The amount preferable range of the electron transfer agent phosphate derivative that exists is maximum 11%, and more preferably 1% to 11%, most preferably 1% to 3%.
The complex of electron transfer agent phosphate derivative
When other character of needs (as, the stability of raising or delivery capability) time, can also use electron transfer agent phosphate derivative complex.This complex is the product of one or more electron transfer agent phosphate derivatives and one or more chelating agent, described chelating agent is selected from amphoteric surfactant, cationic surfactant, has nitrogen functional group's aminoacid and is rich in these amino acid whose protein, as disclosed in international patent publications No.WO 02/40034, it is incorporated by reference this paper.
Preferred chelating agent is selected from aminoacid (as arginine and lysine) and replaces amine suc as formula those uncles of (II):
NR
7R
8R
9
(II)
Wherein
R
7Be selected from the optional C that is interrupted by carbonyl
6-22Alkyl; And
R
8And R
9Independently be selected from H, CH
2COOX, CH
2CHOHCH
2SO
3X, CH
2CHOHCH
2OPO
3X, CH
2CH
2COOX, CH
2COOX, CH
2CH
2CHOHCH
2SO
3X or CH
2CH
2CHOHCH
2OPO
3X, wherein X is H, Na, K or alkanolamine,
Prerequisite is R
8And R
9Be not H simultaneously, and work as R
7When being RCO, R
8Be NCH
3And R
9Be (CH
2CH
2) N (C
2H
4OH)-H
2CHOPO
3Perhaps R
8And R
9Form N (CH together
2)
2N (C
2H
4OH) CH
2COO.
Preferred chelating agent comprises arginine, lysine or lauryl imido grpup dipropionic acid, complexation wherein takes place to form stable complex between basic nitrogen center and phosphate ester.
Term " C
6-22Alkyl " refer to have straight chain, side chain or the cyclic hydrocarbon group of 6 to 22 carbon atoms.Example comprises hexyl, cyclohexyl, decyl, lauryl, tridecyl, myristyl, pentadecyl, cetyl, heptadecyl and octadecyl.
Bioactive compound
Term " bioactive compound " refers to have biological effect in the human or animal and the chemical compound that is used for medical treatment, herding or cosmetic applications.Bioactive compound comprises the medicine or derivatives thereof, particularly its phosphoric acid derivatives.Medicine comprises vitamin, phytochemicals, enamel, dietetic product, peptide, polypeptide, protein or nucleic acid.It is a kind of to should be understood that some bioactive compounds can be classified into surpassing of these kinds apoplexy due to endogenous wind.
The pharmaceutically acceptable salt and the derivant that should be understood that said medicine are included in the scope of the present invention.
Preferably, the scope of the amount of bioactive compound is maximum 5%, and more preferably 0.5 to 3%, most preferably 0.5 to 2%.
Vesicle
When existing, the vesicle diameter range is 50nm to 10,000nm, more preferably 100nm to 500nm, most preferably 300nm to 500nm.
Bioactive compound can be wrapped up by vesicle to small part.
The administration type
Preparation comprises those suitable parenteral, enteral, oral, local, percutaneous, through the preparation of eye, rectum, vagina, intranasal and feeding drug into pulmones.Preparation can be forms such as liquid agent, solution, suspending agent, cream, ointment, lotion, gel, powder, aerosol, paster, enteric coated tablet, capsule, suppository, vaginal suppository or tampon, and can be by the known any method preparation of pharmaceutical field, as Remington JP, The Science and Practice of Pharmacy, AR Gennaro edits, and the 20th edition, Lippincott, describe among the Williams and Wilkins Baltimore, Md (2000).These methods comprise bioactive compound and carrier are formed related step, and if desired, then preparation made the product of wanting.
Preparation can dosage form, preparation or via suitable doser or implants by injection, infusion or implantation (intravenous, subcutaneous etc.) parenteral, described implants contains conventional nontoxic pharmaceutical acceptable carrier and adjuvant.
The preparation that is used for the parenteral use can exist by unit dosage form (for example, cuing open with the single dose peace), perhaps is present in the phial that contains several dosage, wherein can add suitable preservatives.Preparation can be solution, suspending agent, Emulsion, infusion device or the doser that is used to implant form, and perhaps it can be used as the dried powder existence, and water or other suitable carriers dissolve again before use.Except bioactive compound, preparation can comprise that suitable parenteral can accept carrier and/or excipient.Bioactive compound can add microsphere, microcapsule, nanoparticle, liposome etc. and be used for sustained release.And preparation can comprise suspending agent, solubilizing agent, the agent of stabilizing agent pH regulator and/or dispersant.
As mentioned above, preparation can be for being fit to the form of aseptic injection.For preparing such preparation, bioactive compound dissolving or be suspended in the acceptable liquid-carrier of parenteral.Operablely accept carrier and solvent comprises: water; Be adjusted to the water of suitable pH by hydrochloric acid, sodium hydroxide or the suitable buffer agent that adds appropriate amount; 1,3 butylene glycol, Ringers solution; And isotonic sodium chlorrde solution.Aqueous compositions can also contain one or more antiseptic (for example, methyl parahydroxybenzoate, ethylparaben or P-hydroxybenzoic acid n-propyl).Only slightly molten or when being slightly soluble in water when a kind of described chemical compound, can add dissolution enhancers or solubilizing agent, perhaps described solvent can comprise third rare or glycol of 10%-60%w/w etc.
The form of controlled release parenteral compositions can be: aqueous suspension agent, microspheres agent, microcapsule, magnetic microspheres, oil preparation, oil-suspending agent or Emulsion.Perhaps, bioactive compound can be added into biological compatibility carrier, liposome, nanoparticle, implants or infusion device.
For example, the material that is used to prepare microspheres agent and/or microcapsule is that biodegradable/biology can lose the polymer of separating, as poly-acetic acid lactic polyester, poly-(alpha-cyanoacrylate isobutyl), poly-(2-hydroxyethyl-L-glutaminate (glutamnine)) with gather (lactic acid).
Operable biological compatibility carrier is saccharide (for example, glucosan), protein (for example, albumin), lipoprotein or antibody when preparation controlled release parenteral formulation.
The material that is used for implants can right and wrong biodegradable (for example, polydimethylsiloxane) or biodegradable (for example, poly-(caprolactone), poly-(lactic acid), poly-(glycolic) or poly-(ortho esters)).
The preparation that is fit to oral administration conventionally can be used as following form and exists: separate unit, as capsule, cachet or tablet, it can contain the bioactive compound of scheduled volume separately; Powder or granule; Solution, suspending agent or Emulsion.Bioactive substance also can be used as pill, electuary or paste and exists.The tablet and the capsule that are used for oral administration can contain conventional excipients, as bonding agent, filler, wetting agent, disintegrating agent or wetting agent.Tablet can be according to method coating well known in the art.For example, oral liquid can be water or oil suspension, solution, emulsion, syrup or elixir, perhaps can be used as dried powder and exists, and water or other suitable carriers dissolve again before use for they.Such liquid preparation can contain conventional the interpolation, as suspending agent, emulsifying agent, nonaqueous carrier (it can comprise edible oil) or antiseptic.
Be the transdermal topical administration, bioactive compound can be made into ointment, cream or lotion, perhaps as transdermal patch.For example, preparation at the bottom of ointment and cream available water or the oil base, and add suitable thickening agent and/or gellant.Preparation at the bottom of lotion available water or the oil base, and generally also contain one or more emulsifying agents, stabilizing agent, dispersant, suspending agent, thickening agent or coloring agent.
Be suitable for that the preparation of topical comprises in the mouth: lozenge, it is contained in the reactive compound of seasoning substrate, and described seasoning substrate is generally sucrose and arabic gum or gum tragacanth; Pastille, it is contained in the active component in the inertia substrate, described inertia substrate such as gelatin or sucrose and arabic gum; And collutory, it is contained in the active component of suitable liquid-carrier.
The preparation that is suitable for rectally can be used as suppository and exists.Suitable vehicle comprises in cocoa butter and the prior art other materials that generally uses, and suppository can by with bioactive compound with soften or the melt carrier mixes mutually and subsequently by putting cold and mould prepares.
The preparation that is suitable for vagina administration can be used as vaginal suppository, tampon, cream, gel, paste, foam or spray and exists, and except bioactive compound, it contains the known suitable vehicle of prior art.
Be intranasal or feeding drug into pulmones, preparation can solution or suspending agent or as the form administration of dried powder.
Solution and suspending agent generally are moisture (for example, aseptic or pyrogen-free water) and the acceptable cosolvent of physiology (for example, ethanol, propylene glycol or Polyethylene Glycol such as PEG 400).
Such solvent or suspending agent can be chosen wantonly and contain other excipient, for example: antiseptic (as benzalkonium chloride), solubilizing agent or surfactant such as polysorbate are (for example, Tween 80, Span 80, benzalkonium chloride), buffer agent, isoosmotic adjusting agent (for example, sodium chloride), absorption enhancer and viscosifier.Suspending agent also can contain suspending agent (for example, microcrystalline Cellulose, sodium carboxymethyl cellulose).
Solution or suspending agent can directly be applied to nasal cavity by conventional methods, for example use dropper, pipet or spraying.Preparation can single dose or the multiple dose form provide.Back one situation means need provide dosage measuring.In the situation of dropper or pipet, can realize dosage measuring by solution or the suspension of using suitable predetermined.In the spraying situation, for example, can realize by metering property atomisation pump.
Can also realize administration by aerosol preparations to respiratory tract, wherein bioactive compound provides with suitable propellant in compression wrap, described propellant such as Chlorofluorocarbons (CFC), for example, dichlorofluoromethane, Arcton 11 or dichlorotetra-fluoroethane, carbon dioxide or other suitable gas.The aerosol routine also can contain surfactant (as lecithin).Can control drug dose by metering valve is provided.
Perhaps, can provide described chemical compound by dry powder form, the mixture of powders of chemical compound in suitable powder base for example, described powder base such as lactose, starch, starch derivatives (as hydroxypropyl emthylcellulose) and polyvinylpyrrolidine (PVP).Dust carrier will conveniently form gel in nasal cavity.Powder composition can exist by unit dosage form, and for example in the capsule or cartridge case or medicated bag of for example gelatin, powder can be from it by inhaler (as Diskhaler, the trade mark of GlaxoSmithKline) or the administration of dosing aerosol inhaler.
Other excipient
One skilled in the art will know which other excipient can be included in the preparation.The selection of other excipient will be depended on the character of bioactive compound and the form of medication of use.The example of other excipient comprises solvent, thickening agent or gellant, surfactant, buffer agent, lubricant, sweeting agent, disintegrating agent, flavoring agent, coloring agent, antiseptic, aromatic, electrolyte, film forming polymer etc.Suitable sweeting agent comprises sucrose, lactose, glucose, aspartame or glucide.Suitable disintegrating agent comprises corn starch, starch, methylcellulose, polyvinylpyrrolidone, xanthan gum, bentonite, alginic acid or agar.The proper flavor agent comprises Oleum menthae, wintergreen oil, Fructus Pruni pseudocerasi, Fructus Citri junoris or raspberry flavoring agent.Suitable preservatives comprises sodium, benzoate, vitamin E, alpha-tocopherol, ascorbic acid, methyl parahydroxybenzoate, propyl p-hydroxybenzoate or sodium sulfite.
The typical excipient of preparation of the present invention comprises gellant such as carbomer (Carbopol), and it is carboxyl ethylene polymer, antiseptic (as methyl parahydroxybenzoate, butyl p-hydroxybenzoate, ethylparaben, propyl p-hydroxybenzoate and sodium benzoate) and buffer agent (as sodium hydroxide).Excipient can about at most 5% amount exist.
The method for preparing carrier or preparation
The method for preparing carrier comprises electron transfer agent phosphate derivative or its chelating agent is combined with pure, adds water then.Then, prepare preparation by adding bioactive compound to this carrier in any step of the process for preparing carrier.
Usually, alcohol is heated to 55 ℃ or higher temperature, and the electron transfer agent phosphate derivative is dissolved in this alcohol.If bioactive compound is dissolved in alcohol, then when merging with alcohol, the electron transfer agent phosphate derivative adds this bioactive compound, and the watering balance said preparation.
Can in any step of described method, (usually after adding water) add other excipient, as gellant, antiseptic and buffer agent.
Can use any known hybrid technology (as, for example vibrations or vortex) component of carrier and preparation is mixed.
Description of drawings
Embodiment will be described with reference to the drawings, in the described accompanying drawing:
Fig. 1 is the figure that shows mean P TH concentration in the Mus blood plasma.
Fig. 2 is the figure that is presented at the increased radioactivity in the Mus organ behind the local application transdermal TPM-I125-insulin.
Fig. 3 is the figure of insulin level in the Mus blood plasma.
Fig. 4 is the mean change through transdermal insulin (Lispro) treatment back blood sugar concentration.
The specific embodiment
Embodiment
Now, will various embodiments of the present invention/aspect be described with reference to following non-limiting example.
Present embodiment has been investigated human parathyroid hormone (segment 1-34) Transdermal absorption (PTH) of using preparation of the present invention.
Materials and methods
Be prepared as follows test formulation.All percentage ratios are w/w.
| Composition | TPM-01/PTH | TPM-02/PTH |
| PTH-(1-34) (U.S.'s peptide, USA) | 0.1% | 0.1% |
| Contain 2: 1 percentage of T P: T 2The sour form mixture of the phosphorylation tocopherol of P.TP refers to the phosplate of alpha-tocopherol, and T 2P refers to two-tocopherol phosphate ester. | 1% | 1% |
| Ethanol | - | 20% |
| Carbomer | 0.4% | 0.75% |
| Methyl parahydroxybenzoate | 0.1% | 0.1% |
| Water | To 100% | To 100% |
The TPM-02/PTH preparation is the soliquid that seems milk.This shows and has generated vesicle.
Treatment
Sprague-Dawley rat (10-12 week big, male) be divided at random the treatment group (group 1 and group 2, n=6), and stable breeding in independent box to prevent that its Mus that lives together from licking preparation from its back.
The treatment group:
Group 1-100mg TPM-01/PTH/200g body weight, continues 24 hours at every day twice.
Group 2-100mg TPM-02/PTH/200g body weight, continues 24 hours at every day twice.
With Mus anesthesia, weigh and be close to the zone that scrapes 5 * 4cm under the neck.Rat afterbody when anesthesia is got blood, collects blood plasma to measure the PTH concentration before treatment begins.Beginning in second day, for every rat takes by weighing the preparation of suitable dosage, and the finger that uses the band glove is rubbed skin into rat with preparation.In 24 hours, be applied to organize 1 and group 2 with preparation every day twice (morning and evening).The treatment phase is passed through CO when finishing
2Suffocate and kill rat and pass through heart puncturing extracting blood.
PTH analyzes in the blood plasma: by centrifugal from the blood of collecting separated plasma, and be stored in-20 ℃ until analysis.(Immunotopics Inc. USA) comes analyzing rat blood plasma PTH concentration according to manufacturer's description to end user's biological activity PTH1-34 ELISA test kit.
The result
The mean P TH concentration that detects in the blood plasma is summarized in Fig. 1.
Twice TPM-01/PTH used and caused the significantly (p＜0.05*) increase of blood plasma PTH content after 24 hours every day.With respect to the foundation level of not treating rat, average blood plasma PTH concentration has increased 685pg/ml.
Twice TPM-02/PTH used and caused the significantly (p＜0.05*) increase of blood plasma PTH content after 24 hours every day.This increase (1185pg/ml) accounts for 7.7% of accumulated dose, has improved 70% than TPM-01/PTH preparation.
* the result of student ' s t-check
Use the skin-penetrating therapeutic of TPM-02/PTH to improve the PTH concentration in the rat plasma, show that TPM can realize the Transdermal absorption of PTH-(1-34) in 24 hours, significantly improved blood plasma PTH concentration than not treating contrast.Treat after 72 hours, the PTH plasma concentration returns to foundation level.
That has reported studies show that, behind subcutaneous injection therapeutic dose (25 μ g/kg body weight), the PTH concentration in the rat plasma reached peak value in the time of injection ~ 40-60 minute, and metabolism fully in 4 hours.Although be topical application, the rat in this research has received much bigger dosage (500 μ g/kg body weight).Because fast PTH metabolic rate, legitimate inference: the high concentration PTH that kept 24 hours after the initial application only accounts for the little percentage ratio that receives accumulated dose.Therefore, the effective dose that produces of topical application TPM preparation will be more much bigger than the PTH concentration of measuring after 24 hours.
Conclusion
Although TPM-01 and TPM-02 preparation all are effectively sending on the PTH, the PTH that TPM-02/PTH is delivered to blood circulation Duos 70% than TPM-01/PTH.The more effective delivering active ingredients of preparation of the present invention sees through skin and enters systemic circulation.
This method has been described ubiquinone (CoQ)/tocopherol phosphate mixture preparation of preparation 100ml, is used for subsequently the application at preparation of the present invention.Final preparation contains 0.5%CoQ10,1%TPM, 10% ethanol, 1% carbomer, 0.1% methyl parahydroxybenzoate, QS MilliQ.
Equipment and material
Ethanol (AR level)
MilliQ water
Ubiquinone (Kaneka)
Tocopherol phosphate mixture (TPM), containing proportional is tocopherol phosphate ester (TP) and the two-tocopherol phosphate ester (T2P) (Phosphagenics Ltd) of 2: 1 w/w.
Carbomer 934 P USP powder (Croda Surfactants Ltd)
Methyl parahydroxybenzoate BP powder powder (Bronson﹠amp; Jacobs)
1M sodium hydroxide (NaOH)
Balance (Mettler AE 240)
The Falcon pipe
100ml plastic sample container
70 ℃ of water-baths
The multi-vortex spigot
Method
1. accurately take by weighing 0.5g CoQ and go into 50ml Falcon pipe.
2. accurately take by weighing 1g TPM and go into 50ml Falcon pipe.
3. take by weighing 10g (non-ml) ethanol and go into pipe.Closely capping and mixing.
4. in 70 ℃ of water, heat, to help dissolving/fusing component.Per a few minutes manually vibrate once, until CoQ and TPM dissolving.Be heated to the temperature that needs.Be in the CoQ of this concentration, promptly come out from ethanol precipitation once cooling.
5. measure 80ml MilliQ and go into the 100ml shuttle.Closely capping and place water-bath 5 minutes to add hot water.
6. the CoQ/TPM/ alcoholic solution with heating is poured directly into MilliQ.
7. add a cover immediately and manually thermal agitation with blending ingredients.Said preparation has opaque yellow appearance.Vortex 5 minutes.
8. accurately take by weighing the 1g carbomer and the 100mg methyl parahydroxybenzoate is gone into weighing plate.Slowly be added into CoQ/TPM solution, violent vortex between each the interpolation.70 ℃ of water-bath short-term heated sample to help sample dissolution.
9. after having added all carbomer/methyl parahydroxybenzoate, vortex reaches even denseness until component, but should the stage also not form gel.
10. add 3ml 1M NaOH, add a cover and thermal agitation.
11., then detect pH if preparation has formed gel.Carbomer will form the best gel of pH7-8.
12. repeating step 10 and 11, the denseness of wanting until formation.
13. if desired, be supplemented to 100g with MilliQ.
14. vortex is 5 minutes once more.
15. container is wrapped in the aluminium foil to prevent the photodissociation of CoQ.
16. by second day, any undissolved carbomer absorbed the moisture in the preparation and has formed clarifying gel pockets (pockets of gel).Thermal agitation forms even denseness until preparation.
Preparation is the soliquid that seems milk.This shows that vesicle forms.
Present embodiment is investigated
Present embodiment has been investigated the Transdermal absorption of using the coenzyme Q10 (CoQ10) of preparation of the present invention.
Materials and methods
Ethanol (AR level)
MilliQ water (inner supply)
Tocopherol phosphate mixture (TPM), containing proportional is 2: the tocopherol phosphate ester (TP) of 1w/w and two-tocopherol phosphate ester (T2P) (Phosphagenics Ltd).
Ubiquinone (CoQ) (Kaneka, Japan)
Nivea Visage anti-wrinkle Q10 day care (Beiersdorf)
Carbomer 934 P USP powder (Croda Surfactants Ltd)
Methyl parahydroxybenzoate BP powder (Bronson﹠amp; Jacobs)
1M sodium hydroxide (NaOH)
Balance (Mettler AE 240)
The Falcon pipe
100ml plastic sample container
55 ℃ of water-baths
The multi-vortex spigot
Test formulation
CoQ contrast: the CoQ10 amount that can use the CoQ control formulation to be evaluated to pass skin when not containing TPM.Form: (Kaneka, Japan), 10% ethanol, 1% carbomer, 0.1% methyl parahydroxybenzoate, water is supplemented to 100% to 0.5%CoQ10.
Take by weighing 0.5g CoQ10 and go into 50ml Falcon pipe.Use platform balance to add 10g ethanol.Closely sealing cap and mixing.In 55 ℃ of water-baths, heat, to help dissolving/fusing CoQ.Be heated to the temperature that needs.Be in the CoQ of this concentration, promptly come out from ethanol precipitation once cooling.Measure 80ml water and go into the 100ml shuttle.The CoQ/ alcoholic solution of heating is poured directly in the water.Add a cover immediately and manually thermal agitation with blending ingredients.Vortex 5 minutes.Number of C oQ10 comes out from solution precipitation, and oily is orange around container.Because the insoluble character of CoQ10, this is inevitable.Accurately take by weighing the 1g carbomer and the 100mg methyl parahydroxybenzoate is gone into weighing plate.Pour preparation and vortex into, until reaching even denseness, but should the stage also not form gel.Add 3ml 1M NaOH, add a cover and thermal agitation.If preparation has formed gel, then detect pH.Carbomer will form the best gel of pH 7-8.Repeat to add 3ml 1M NaOH, vibration and detect pH, until gel formation.If desired, be supplemented to 100g with MilliQ.Vortex is 5 minutes once more.Container is wrapped in the aluminium foil to prevent the photodissociation of CoQ.By second day, any undissolved carbomer absorbed the moisture in the preparation and has formed clarifying gel pockets.Violent vortex forms even denseness until preparation.
TPM contrast: use the TPM control formulation to measure the influence of TPM to endogenous CoQ10 concentration.Form: 1%TPM, 10% ethanol, 1% carbomer, 0.1% methyl parahydroxybenzoate, water is supplemented to 100%.Do not contain CoQ10 in the said preparation.
Take by weighing 1g TPM and go into 50ml Falcon pipe.Use platform balance to add 10g ethanol.Closely sealing cap and mixing.In 55 ℃ of water-baths, heat, to help dissolving/fusing TPM.Be heated to the temperature that needs.Measure 80ml water and go into the 100ml shuttle.The TPM/ alcoholic solution of heating is poured directly in the water.Preparation obtains emulsus character immediately.Add a cover immediately and manually thermal agitation with blending ingredients.Vortex 5 minutes.Accurately take by weighing the 1g carbomer and the 100mg methyl parahydroxybenzoate is gone into weighing plate.Pour preparation and vortex into, until reaching even denseness, but should the stage also not form gel.Add 3ml1M NaOH, add a cover and thermal agitation.If preparation has formed gel, then detect pH.Carbomer will form the best gel of pH7-8.Repeat to add 3ml 1M NaOH, vibration and detect pH, until gel formation.If desired, water is supplemented to 100g.Vortex is 5 minutes once more.Container is wrapped in the aluminium foil to prevent the photodissociation of CoQ.By second day, any undissolved carbomer absorbed the moisture in the preparation and has formed clarifying gel pockets.Violent vortex forms even denseness until preparation.
TPM-02/CoQ: as above embodiment 2 is described, prepares TPM-02/CoQ of the present invention.Form: 0.5%CoQ10,1%TPM, 10% ethanol, 1% carbomer, 0.1% methyl parahydroxybenzoate, water is supplemented to 100%.
Nivea Visage anti-wrinkle Q10 day care (Beiersdoff, Germany): but NiveaVisage is the facial cream that commercial sources obtains, and propagates the source into effective CoQ10, is used for skin.Because known accurate CoQ10 content, Nivea Visage is suitable with TPM-02/CoQ on weight.Form: the unknown.
The treatment group
This pula-Dao comes (family name) rat (10-12 week is big, male) available from Animal Services, Monash University, and treating beginning prospective adaptation department's Animal House (DepartmentalAnimal House) at least 5 days.Animal is divided into treatment (n=6) at random, and is housed in the single box with the Mus that prevents to live together and licks preparation from its back.Food (the rat experiment chamber slice foodstuffs of standard freely is provided; Barastoc, Australia) and water.
Group 1-does not treat
Group 2-100mg CoQ contrast/200g body weight, continues 24 hours at every day twice
Group 3-100mg TPM contrast/200g body weight, continues 24 hours at every day twice
Group 4-100mg TPM-02/CoQ/200g body weight, continues 24 hours at every day twice
Group 5-100mg Nivea Visage frost/200g body weight, continues 24 hours at every day twice
Group 6-100mg CoQ contrast/200g body weight, continues 48 hours at every day twice
Group 7-100mg TPM contrast/200g body weight, continues 48 hours at every day twice
Group 8-100mg TPM-02/CoQ/200g body weight, continues 48 hours at every day twice
Group 9-100mg Nivea Visage cream /200g body weight, continues 48 hours at every day twice
With Mus anesthesia, weigh and be close to the zone that scrapes 5 * 4cm under the neck.Beginning in second day, for every rat takes by weighing the preparation of appropriate amount, and the finger that uses the band glove rubs skin into rat with preparation, twice of every day (morning and evening), continues 24 or 48 hours.Preparation is limited to rat impalpable skin of back zone of resonable when hair.
The analysis of CoQ10 in skin and the blood plasma: when the treatment phase finishes, kill rat by using CO2 gas to suffocate.By heart puncturing extracting blood, packing into has added in the collecting pipe of heparin, and centrifugal separation plasma.With the skin area of the abundant cleaning scraper hair of distilled water, with remove residual from the teeth outwards and any CoQ10 of absorption in the zone of exsomatizing.Basically according to the method for people such as Aber ((1992) Distribution andredox state of ubiquinones in rat and human tissues.Arch BiochemBiophys 295:230-234), it is quantitative from the extraction and the HPLC of tissue to carry out CoQ.
Statistical analysis: the result is expressed as meansigma methods ± SD.Carry out Student ' s t-check to determine whether the CoQ concentration from blood plasma and skin extraction exists significant difference between the treatment group.
The result
Average CoQ10 concentration in Table I-treatment back blood plasma and the skin
| Treatment | Average CoQ in the blood plasma 10(ng/ml) | Average CoQ in the skin 10(μg/g) | ||
| 24 hours | 48 hours | 24 hours | 48 hours | |
| Do not treat CoQ contrast TPM contrast TPM-02/CoQ Nivea Visage | 27.67±4.97 35.00±6.45 33.67±7.06 59.33±16.47 41.33±4.59 | - 36.33±4.84 28.33±5.79 42 33±8.80 29.83±6.18 | 0.24±0.04 0.74±0.20 0.32±0.02 6.13±0.98 0.38±0.05 | - 1.73±0.27 0.61±0.12 10.59±4.08 0.62±0.11 |
Blood plasma
TPM-02/CoQ is to the rat back zone in twice application every day, causes significantly (p＜0.05) increase (Table I) of CoQ10 content in the blood plasma.Use the average blood plasma CoQ10 concentration after TPM-02/CoQ treats to increase by 114% (p＜0.05) with respect to observed endogenous CoQ10 concentration during treatment does not contrast.On the contrary, CoQ contrast and TPM contrast only can improve average blood plasma CoQ10 concentration 26% and 22% respectively.Latter two increases neither one and has reached the significance on the statistics.Importantly compare with respect to the CoQ control formulation that lacks TPM, TPM-02/CoQ significantly (p＜0.05) has improved plasma C oQ10 concentration 70%, proves that TPM and ethanol directly improve the Transdermal absorption of CoQ10.
Not treatment contrast relatively, after the odd-numbered day treatment, Nivea Visage makes plasma C oQ10 concentration improve 49%.Yet the plasma C oQ10 amount that produces by Nivea Visage significantly is less than (44%; P＜0.05) the plasma C oQ10 amount that produces by the TPM-02/CoQ treatment.
Skin
Use the TPM-02/CoQ treatment beginning make in 24 hours in the skin endogenous CoQ10 concentration significantly (p＜0.05) improved 2454% (Table I).By 48 hours, this raising rose to 4312% of endogenous concentration.On the contrary, CoQ contrast and TPM contrast make mean skin CoQ10 concentration beginning to have improved 208% and 33% respectively in 24 hours, have improved 621% and 154% in 48 hours.Though be significant (p＜0.05), the value that uses TPM contrast treatment back to increase seems and almost is no advantage.With respect to the CoQ contrast, TPM-02/CoQ makes mean skin CoQ10 improve 728% and 512% respectively.
TPM-02/CoQ remarkable (p＜0.05) after 24 hours has improved (1513%) mean skin CoQ10 concentration, and Nivea Visage fails to produce the skin CoQ10 concentration that significantly improves that is higher than other control formulation.
Conclusion
TPM/ ethanol preparation has strengthened the dissolubility of CoQ10 and Transdermal absorption subsequently, compares with control formulation (but comprising the beauty treatment CoQ10 source that commercial sources obtains), has significantly improved the CoQ10 concentration of blood plasma and skin.For known topical application and absorption, use TPM/ ethanol preparation preparation chemical compound of the present invention to have great potential with molecule of poor oral administration biaavailability, skin specificity or adverse side effect (in digestion process, manifesting).
Preparation contains the preparation of insulin as mentioned above.Preparation is composed as follows:
| Composition | The TPM-02/ insulin |
| Insulin | 60 units/g gel |
| Contain 2: 1 percentage of T P: T 2Phosphorylation tocopherol (TPM) mixture of P.TP refers to the phosplate and the T of alpha-tocopherol 2P refers to two-tocopherol phosphate ester. | 2 |
| Ethanol | |
| 30% | |
| Carbomer 934 | 1% |
| Water | To 100% |
Present embodiment has been investigated the transdermal administration of the insulin that uses the TPM preparation
| Formulation components | LISPRO, human insulin analogue (Eli Lilly) bovine insulin (Sigma) 3-[ 125I] iodo tyrosyl A14) insulin, people's recombinant (AmershamBiosciences, code IM166, lot number B0602) ( 125The I-insulin) the blended alpha-tocopherol phosphate ester of TPM-(TP) and two-tocopherol phosphate ester (T2P; 2: 1) |
| The dosage particles component | 32.5U LISPRO/kg body weight 10U bovine insulin/kg body weight
125I-insulin (people's recombinant; 400nCi)/ |
| Experiment | |
| 1 to 3The animal |
Sprague-Dawley rat sex: male weight range: 220-450g age: 10-12 week pig |
Carry out four independently experiments, independently proved the insulin transdermal delivery behind the use TPM preparation topical.Use bovine insulin, Semilente Insulin analog (LISPRO) or radiolabeled biosynthetic human insulin to prepare TPM.Reduce by the blood sugar concentration behind the raising of plasma insulin concentration, subcutaneous radiological measuring or the glucose load and to estimate successful transdermal administration.
Experiment 1: improve plasma insulin concentration
Testing the previous day, under the slight anesthesia (ether) hair is being scraped in the skin of back zone of male sprague-Dawley rat (220-300g).When sleeping soundly, the rat of weighing is to calculate each rat pentobarbital that needs and the dosage for the treatment of preparation.The rat overnight fasting (~ 16h), and freely supply water.
Rat back anesthesia in the morning, and in experimentation, keep anesthesia.Test formulation contains 2%TPM, bovine insulin (3U/100 μ l; Sigma), ethanol (30%) and carbomer (1%), water is supplied.Rat has received the final insulin dose of 10U/kg body weight.Matched group has received and has contained TPM but the same preparation of insulin-containing not.The finger of common use band glove will contrast (n=2) and TPM-insulin preparation (n=2) is used or rubbed into skin.1,2,3,4 and 6 hour collection blood plasma after the administration.
Use specificity to measure the amount of the insulin that exists in the plasma sample at the competition radioimmunoassay (Linco Research Inc.) of bovine insulin.
Experiment 2: use radioactive probe to detect the Transdermal absorption of insulin
Prepare to be used for the sprague-Dawley rat (300-450g) of preparation topical according to experiment 1.The rat overnight fasting (~ 16h), and freely supply water.
To contain radiolabeled biosynthetic human insulin (
125The I-insulin, Amersham Biosciences) form gel (TpM-with 2%TPM, 30% ethanol, 1% carbomer and water preparation
125The I-insulin).Dosage local application TPM-with ~ 400nCi/ Mus (n=4)
125I-insulin (as above).Contrast Mus (n=5) receives the preparation that does not contain TPM, to illustrate the effect of TPM in Transdermal absorption.Stable breeding separately after the rat administration, and freely supply food and water.After 5 hours, put to death rat and also shift out organ, weigh and place scintillation vial to measure each organ radioactivity total amount.Clean skin to remove any unabsorbed I that remains in skin surface
125-insulin.
Experiment 3: use transdermal insulin blood sugar lowering
Prepare to be used for the sprague-Dawley rat (220-300g) of preparation topical according to experiment 1.The rat overnight fasting (~ 16h), and freely supply water.
Quick-acting human insulin analogue LISPRO (Eli Lilly) prepare with 2%TPM, 30% ethanol, 1% carbomer and water and form gel (TPM-LISPRO).Treatment group (n=15) received the topical therapeutic 30 minutes of TPM-LISPRO (dosage is 32.5U LISPRO/kg body weight) before glucose load, so that LISPRO enters systemic circulation if having time.Matched group (n=15) receives the preparation of no LISPRO.Glucose (30%w/v) is with the dosage IP injection of 2g/kg body weight (2ml/300g Mus).
Rat keeps anesthesia (pentobarbital) in whole experiment, use Medisense Optium blood sugar monitoring instrument (Abbott) to measure blood glucose from afterbody.In 5 minutes and measuring blood concentration after 5 minutes again behind the glucose load.Then, per 10 minutes measuring blood, lasting ~ 2-2.5 hour.The instant blood sugar concentration of measuring before the glucose load is deducted from all subsequent values, to determine the change of blood sugar of every rat in the experiment.Calculate the change of blood sugar meansigma methods of each time point and make curve (Fig. 3).As the non-diabetic rat of using in this research, judge that the effectiveness of TPM-LISPRO has reduced the blood sugar concentration peak value for comparison according to animal.
Calculate the area under curve of each Mus, and use relatively group of Student ' s t-check.
Experiment 4: use transdermal insulin blood sugar lowering
Before research, prepare eight pigs at least 5 days, have the conduit that two surgeries are prepared.Pig is trained in approximately 3:00pm feed, so that it adapts to overnight fast.Experimental design is the single counter-rotating with two treatments, and described two treatments are to contain the TPM-02 gel of insulin or the TPM-02 gel of insulin-containing not.Vein (IV) infusion at least 1 day at interval.On infusion same day, pig was got blood in per 15 minutes, continue 1 hour, to obtain to use the basic blood sugar concentration before the gel.After 30 minutes, beginning infusion glucose (0.33g/kg/h) and xylazine (0.033mg/kg/h), blood sampling continues other 3 hours.Use glucometer analyzing blood glucose immediately.During studying, the conduit in pig stops up, and therefore has only 7 pigs to study.And, getting the blood same day (insulinize), the sampling catheter in pig stops up.Therefore, contrast pig and insulinize pig gets blood natural law respectively not 7 and 6.
Use REML to analyze blood glucose level data, fixed effect comprises treatment (contrast or insulin) and gets the blood time, and stochastic model comprises pig and gets blood day.In addition, the blood glucose to pretreat phase and last 2 and 4 samples averages.Also use REML to analyze these data, fixed effect is for getting the blood time (before or after using gel and infusion), and stochastic model comprises pig and gets blood day.For the analysis of back, data transform through logarithm.
Result and discussion
Preliminary experiment 1: improve plasma insulin concentration
In tentative experiment, the topical application of TPM-insulin can improve serum insulin concentration (Fig. 3).In being controlled animal, the treatment that is increased in of serum insulin concentration reached peak value in back 4 hours.Insulin concentration in the control animal is in this decline in period or fail to reach similar concentration for the treatment of animal.It is few to be used for this preliminary experiment animal number, can not carry out statistical evaluation; Yet the positive trend of successful Transdermal absorption is conspicuous, has guaranteed the further investigation in the greater amount experiment.
Experiment 2: use radioactive probe to detect the insulin Transdermal absorption
Obtained the positive evidence that serum insulin concentration increases in tentative experiment, we seek the Transdermal absorption that concluding ground proof is used the TPM insulin.For this reason, we are with radiolabeled insulin form preparation TPM, and the purpose of using radioactive decay is Transdermal absorption of monitoring " heat " insulin and the distribution (if there is) in Mus subsequently.The result shows that TPM can successfully drive
125The Transdermal absorption of I-insulin (Fig. 2).It is (p＜0.001) that significantly improves that the radioactive level that detects in the skin of application site is compared with control animal.Because the skin surface of every pig is through cleaning, so this radioactivity is present in the deep skin layer.Importantly, just the subcutaneous fat under the application region has the significantly radioactive level of (P＜0.05) raising compared with the control, has proved that concluding TPM drives the ability of insulin Transdermal absorption to lower-hierarchy.
Experiment 3: use transdermal insulin blood sugar lowering
The verified insulin Transdermal absorption of success when preparing with TPM, we seek to check the molecule of being sent whether can effectively enter systemic circulation with blood sugar lowering.The fasting rat carried out the glucose tolerance test in 30 minutes behind topical application TPM-LISPRO, in measuring blood interval (Fig. 4) subsequently.Use in the animal of TPM-LISPRO treatment blood sugar concentration compared with the control significantly (p＜0.02) descend, proved transdermal administration and the activity of the LISPRO that is transported subsequently.Therefore, TPM can transport big bioactive molecule (as insulin) and pass skin.
Experiment 4: use transdermal insulin blood sugar lowering
This research launches on following Mus research basis: but wherein glucose tolerance test shows transdermal insulin preparation passes skin and be biological utilisation.It obtains estimating in pig by the IV glucose tolerance test of using the TPM-02/ insulin.
By using IV dosed administration glucose to replace initial oral glucose test (it does not play predictive role) to improve method.In addition, xylazine (preventing the chemical substance that insulin discharges at pancreas) is total to infusion with glucose.
The general evaluation system effect of TPM-02/ insulin is very significant (p＜0.005).Obvious effects appears at the infusion later stage, and this moment, blood glucose reached high initial value.In high initial value, the blood glucose increase is obviously lower in the pig of reception transdermal insulin preparation, significantly improving in this expression blood glucose contrast.Data show, the insulin Transdermal absorption.
Infusion shows as the good model system of measuring the insulin Transdermal absorption in the time of glucose and insulin secretion inhibitor (as xylazine).Further the work purposes that should expand present model to be paying close attention to the effect of transdermal administration in more unexpected blood glucose increase process, and should prolong search time and how long can continue to measure described sending.Further research can also (that is, the diabetes patient be carried out on) the model system, as streptozocin diabetes pig being suitable for target.That is, the chemicals streptozocin has been treated pig, and streptozocin destroys the insulin secretory cell of pancreas, and this cell makes pig suffer from diabetes.
Conclusion:
The result who is proposed proves that blended tocopherol phosphate ester (TPM) can successfully drive the Transdermal absorption of macromole (as insulin).The intradermal of application site, below subcutaneous fat and blood in confirmed the insulin concentration that increases.Importantly, the glucose tolerance test shows is activated and effective blood sugar lowering by the molecule sent.This is positive discoveries for diabetes, and following hope is provided: can obtain Noninvasive insulin administration method to alleviate a day injection discomfort.
Propose to use vertical proliferation (Franz) cell and skin to experimentize, test the flux rate and the permeability of several formulations distortion from pig and people.This technology will be accelerated the optimization of TPM-insulin preparation.
Prepared as mentioned above and contained atropinic preparation.Said preparation is composed as follows:
| Composition | The TPM-02/ atropine |
| The |
1% |
| Contain 2: 1 percentage of T P: T 2Phosphorylation tocopherol (TPM) mixture of P.TP refers to the phosplate of alpha-tocopherol, and T 2P refers to two-tocopherol phosphate ester. | 2 |
| Ethanol | |
| 30% | |
| Carbomer 934 | 1% |
| Water | To 100% |
To contain 2: the phosplate of the alpha-tocopherol of ratio (TP) and two-tocopherol phosphate ester (T
2P) the TPM vesicle of mixture is exposed to mimic gastric juice and intestinal juice, to determine whether Enteral formulations of the present invention can bear digestive tract environment.
The 2%TPM that use comprises the fluorescent dye rhodamine 6G prepares vesicle, and uses fluorescence activated cell sorting (FACS) to analyze the distribution of vesicle colony.
Prepare mimic gastric juice and intestinal juice according to American Pharmacopeia.Gastric juice is pepsic acid solution, pH1.2.Use the pancreatin powder in the phosphate buffer of pH6.8, to prepare intestinal juice.
Vesicle is exposed to two kinds of liquid respectively.Exposure to simulated gastric fluid produces bigger vesicle and/or vesicle aggregation.Exposure to simulated intestinal fluid does not almost influence the vesicle outward appearance, and it keeps initial distribution of sizes.
As mentioned above, prepare the preparation that comprises it, whether can form vesicle with check TPM complex from the TPM complex.Said preparation is composed as follows:
| Composition | %w/w |
| Contain 2: 1 percentage of T P: T 2The lauryl imido grpup dipropionic acid tocopherol phosphate mixture (TPM) of P.TP refers to the phosplate of alpha-tocopherol, and T 2P refers to two-tocopherol phosphate ester. | 3.2 |
| Ethanol | |
| 30% | |
| Water | To 100% |
Form vesicle according to said preparation.
The word that uses in this description ' comprises ' and ' comprising ' not invention of requirement for restriction protection, comprises any distortion or increase.
To be conspicuous to adjustment of the present invention and improvement to those skilled in the art.Such adjustment and improvement expection are included in the scope of the present invention.
Claims (36)
1. carrier that is used to use bioactive compound, this carrier comprises: one or more C
1-C
4Alcohol, polyhydric alcohol and polymer thereof, water and one or more two-and/or list-(electron transfer agent) phosphate derivative or its complex.
2. according to the carrier of claim 1, this carrier is C
1-C
4Alcohol, polyhydric alcohol and polymer thereof are selected from methanol, ethanol, propanol, isopropyl alcohol, butanols and ethylene glycol or its combination.
3. according to the carrier of claim 2, wherein said C
1-C
4Alcohol is ethanol.
4. according to the carrier of claim 1 or claim 2, wherein said C
1-C
4Alcohol, polyhydric alcohol and polymer thereof exist with 0.5% to 50%, 5% to 40% or 10% to 30% amount.
5. according to the carrier of claim 1, comprising one or more two-(electron transfer agent) phosphate derivatives.
6. according to the carrier of claim 1, this carrier comprises the combination of one or more two-(electron transfer agent) phosphate derivatives and one or more list-(electron transfer agent) phosphate derivatives.
7. according to the carrier of claim 1, wherein said two-and/or list-(electron transfer agent) phosphate derivative be the phosphate ester of electron transfer agent, wherein said phosphate ester is by electron transfer agent two-or the orthophosphate or the pyrophosphate of list-replacement.
8. according to the carrier of claim 1, wherein said two-and/or list-(electron transfer agent) phosphate derivative or its complex be selected from: the hydroxychroman phosphate derivative, be phosphate derivative, hydroxy kind carotene phosphate derivative, calciferol phosphate derivative and the ascorbic acid phosphoric acid esters derivant of the hydroquinone of reduction form electron transfer agent K1 and ubiquinone.
9. carrier according to Claim 8, wherein said hydroxychroman phosphate derivative is selected from α, β, γ and the δ tocol phosphate derivative of enantiomer and racemic form.
10. according to the carrier of claim 9, wherein said tocol phosphate derivative is tocopherol phosphate derivative or tocotrienol phosphate derivative.
11. according to the carrier of claim 10, wherein said tocol phosphate derivative is selected from: two-tocopherol phosphate derivative, two-tocopherol, two-phosphate derivative, two-tocotrienol phosphate derivative, list-tocopherol phosphate derivative, list-tocopherol two-phosphate derivative and list-tocotrienol phosphate derivative.
12. according to the carrier of claim 11, wherein said tocol phosphate derivative is two-tocopherol phosphate derivative.
13. according to the carrier of claim 11, wherein said tocol phosphate derivative is the combination of two-tocopherol phosphate derivative and list-tocopherol phosphate derivative.
14. according to the carrier of claim 13, wherein the ratio of list-tocopherol phosphate ester and two-tocopherol phosphate ester is 4: 1 to 1: 4 or 2: 1.
15. according to the carrier of claim 1, wherein said two-and/or list-(electron transfer agent) derivant and the reaction of one or more chelating agent to generate complex.
16. according to the carrier of claim 15, wherein said chelating agent is selected from: amphoteric surfactant, cationic surfactant, have nitrogen functional group's aminoacid and contain amino acid whose protein with nitrogen functional group
17. according to the carrier of claim 16, wherein said chelating agent has following formula (II):
NR
7R
8R
9
(II)
Wherein,
R
7Be selected from the optional C that is interrupted by carbonyl
6-22Alkyl; And
R
8And R
9Independently be selected from H, CH
2COOX, CH
2CHOHCH
2SO
3X, CH
2CHOHCH
2OPO
3X, CH
2CH
2COOX, CH
2COOX, CH
2CH
2CHOHCH
2SO
3X or CH
2CH
2CHOHCH
2OPO
3X, wherein X is H, Na, K or alkanolamine,
Prerequisite is R
8And R
9Be not H simultaneously, and work as R
7When being RCO, R then
8Be NCH
3And R
9Be (CH
2CH
2) N (C
2H
4OH)-H
2CHOPO
3Perhaps R
8And R
9Form N (CH together
2)
2N (C
2H
4OH) CH
2COO.
18. according to the carrier of claim 17, wherein said chelating agent is arginine, lysine and lauryl imido grpup dipropionic acid.
19. according to the carrier of claim 1, wherein said electron transfer agent phosphate derivative or its complex exist with maximum 11%, 1% to 11% or 1% to 3% amount.
20. according to the carrier of claim 1, wherein said water exists with 50% to 99%, 60% to 95% or 70% to 90% amount.
21. according to the carrier of claim 1, this carrier is the form of vesicle.
22. according to the carrier of claim 21, the diameter of wherein said vesicle is 50nm to 10,000nm, 100nm to 500nm, or 300nm to 500nm.
23. one or more C
1-C
4Alcohol, polyhydric alcohol and polymer thereof, water and one or more two-and/or list-(electron transfer agent) phosphate derivative or its complex be used for using the purposes of the carrier of bioactive compound in production.
24. a method that is used to prepare the carrier of claim 1, this method comprises the steps:
(c) with one or more two-and/or list-(electron transfer agent) phosphate derivative or its complex and one or more C
1-4Alcohol, polyhydric alcohol or its combination of polymers; And
(d) water is added in the combination of step (a).
25. a preparation that comprises bioactive compound and carrier, described carrier comprises one or more C
1-C
4Alcohol, polyhydric alcohol and polymer thereof, water and one or more two-and/or list-(electron transfer agent) phosphate derivative or its complex.
26. according to the preparation of claim 25, wherein said bioactive compound is medicine or its phosphate derivative.
27. according to the preparation of claim 26, wherein said medicine is selected from: vitamin, phytochemicals, enamel, dietetic product, hormone, peptide, polypeptide, protein and nucleic acid.
28. according to the preparation of claim 27, wherein said medicine is selected from: neuroleptics, narcosis analgesic, antiinflammatory, anticarcinogen, hydryllin, antianginal agent, the agent of antilipemic obstacle, antidiabetic and hormone analogs.
29. according to the preparation of claim 28, wherein said medicine is selected from: ubiquinone, human parathyroid hormone, insulin, glucagon like peptide, morphine, oxycodone, elastin, retinol and collagen.
30. according to the preparation of claim 25, the amount of wherein said bioactive compound reaches 5%, 0.5% to 3% or 0.5% to 2%.
31. according to the preparation of claim 30, said preparation further comprises other excipient.
32. according to the preparation of claim 31, wherein said excipient is selected from: solvent, thickening agent or gellant, surfactant, buffer agent, lubricant, sweeting agent, disintegrating agent, flavoring agent, coloring agent, antiseptic, aromatic, electrolyte and film forming polymer.
33. according to the preparation of claim 31, wherein said excipient exists with maximum 5% amount.
34. a method that is used to prepare the preparation of claim 25, this method comprises that described carrier comprises one or more C with bioactive compound and carrier-bound step
1-C
4Alcohol, polyhydric alcohol and polymer thereof, water and one or more two-and/or list-(electron transfer agent) phosphate derivative or its complex.
35. a method that is used to use bioactive compound, this method comprise that described carrier comprises one or more C with described bioactive compound and carrier-bound step
1-C
4Alcohol, polyhydric alcohol and polymer thereof, water and one or more two-and/or list-(electron transfer agent) phosphate derivative or its complex.
36. according to the method for claim 35, wherein said bioactive compound and carrier are by parenteral, enteral, oral, local, percutaneous, use through eye, rectum, vagina, intranasal or feeding drug into pulmones.
Applications Claiming Priority (7)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| AU2005903198 | 2005-06-17 | ||
| AU2005903198A AU2005903198A0 (en) | 2005-06-17 | Transdermal delivery of large molecules | |
| AU2005904737 | 2005-08-30 | ||
| AU2005904737A AU2005904737A0 (en) | 2005-08-30 | Drug formulation | |
| AU2006902726A AU2006902726A0 (en) | 2006-05-19 | Carrier | |
| AU2006902726 | 2006-05-19 | ||
| PCT/AU2006/000839 WO2006133506A1 (en) | 2005-06-17 | 2006-06-16 | A carrier comprising one or more di and/or mono-(electron transfer agent) phosphate derivatives or complexes thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101247834A true CN101247834A (en) | 2008-08-20 |
| CN101247834B CN101247834B (en) | 2013-10-30 |
Family
ID=39947856
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN200680021329.XA Active CN101247834B (en) | 2005-06-17 | 2006-06-16 | Carrier comprising one or more di and/or mono-(electron transfer agent) phosphate derivatives or complexes thereof |
Country Status (2)
| Country | Link |
|---|---|
| CN (1) | CN101247834B (en) |
| ZA (1) | ZA200711235B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI507193B (en) * | 2010-03-30 | 2015-11-11 | Phosphagenics Ltd | Transdermal delivery patch |
| CN108601732A (en) * | 2015-12-09 | 2018-09-28 | 磷肌酸有限公司 | pharmaceutical preparation |
| CN113376186A (en) * | 2021-05-12 | 2021-09-10 | 赵静 | High-precision intelligent erythrocyte folic acid detector and detection method thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2657526B1 (en) * | 1990-01-31 | 1994-10-28 | Lvmh Rech | USE OF AN ALPHA-TOCOPHEROL PHOSPHATE, OR ONE OF ITS DERIVATIVES, FOR THE PREPARATION OF COSMETIC, DERMATOLOGICAL, OR PHARMACEUTICAL COMPOSITIONS; COMPOSITIONS THUS OBTAINED. |
| JP4257479B2 (en) * | 2000-06-30 | 2009-04-22 | ライオン株式会社 | Method for producing dentifrice composition containing ascorbic acid phosphate or salt thereof |
| CN1216061C (en) * | 2001-03-02 | 2005-08-24 | 昭和电工株式会社 | Ascorbic acid 2-phosphate metal salt with low calcium content |
-
2006
- 2006-06-16 CN CN200680021329.XA patent/CN101247834B/en active Active
-
2007
- 2007-12-03 ZA ZA200711235A patent/ZA200711235B/en unknown
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI507193B (en) * | 2010-03-30 | 2015-11-11 | Phosphagenics Ltd | Transdermal delivery patch |
| CN108601732A (en) * | 2015-12-09 | 2018-09-28 | 磷肌酸有限公司 | pharmaceutical preparation |
| CN114712308A (en) * | 2015-12-09 | 2022-07-08 | 磷肌酸有限公司 | Pharmaceutical preparation |
| CN113376186A (en) * | 2021-05-12 | 2021-09-10 | 赵静 | High-precision intelligent erythrocyte folic acid detector and detection method thereof |
| CN113376186B (en) * | 2021-05-12 | 2022-12-23 | 无锡科金生物科技有限公司 | High-precision intelligent erythrocyte folic acid detector and detection method thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN101247834B (en) | 2013-10-30 |
| ZA200711235B (en) | 2008-10-29 |
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