CN1174713C - Microporation method of human skin for drug delivery and monitoring applications - Google Patents

Abstract

A method of enhancing the permeability of the skin (120, 274) to an analytic for diagnostic purposes or to a drug for therapeutic purposes is described utilizing micro-pore and optionally sonic energy and a chemical enhancer. If selected, the sonic energy may be modulated by means of frequency modulation, amplitude modulation, phase modulation, and/or combinations thereof. Micro-pore is accomplished by (a) ablating the stratum corneum (274) by localized rapid heating of water such that water is vaporized, thus eroding cells; (b) puncturing the stratum corneum (274) which a micro-lancet calibrated to form a micro-pore of up to about 1000 m in diameter (c) ablating the stratum corneum (274) by focusing a tightly focused beam of sonic energy onto the stratum corneum (274); (d) hydraulically puncturing the stratum corneum (274) with a high-pressure jet of fluid to form a micro-pore of up to about 1000 m in diameter; or (e) puncturing the stratum corneum (274) with short pulses of electricity to form a micro-pore of up to about 1000 m in diameter.

Description

The human body skin micropunch equipment that is used for loading and detection

Cross-reference with relevant application

The application is the application No.08/520 that submits to August 29 nineteen ninety-five, 547 part continuation application, the latter is again the No.08/152 that submitted on November 15th, 1993,442 applications (existing United States Patent (USP) 5,458,140) and the No.08/152 that submitted in 8th of December in 1993,174 (existing U.S. Patent No.s 5,445,611) part continuation application; The provisional application No.60/008 that the application submits on October also claimed nineteen ninety-five 30,043 interests.

Background of invention

The present invention relates generally to the interior analyte of monitoring body and percutaneous carries medicine to intravital field.More specifically, the present invention relates to by the horny layer micropunch to increase the damaging minimum method of percutaneous permeability to non-damage, this method can be used in combination with acoustic energy, chemosmosis promoter, pressurization etc., optionally promoting analyte outwards to flow out and detect it, or promote drug conveying in body from body.

Horny layer is mainly undertaken known skin barrier effect.Therefore, it is to flowing out the external maximum barrier that provides with the analyte percutaneous in medicine and other molecule percutaneous inflow body.Horny layer (skin outside horn shape layer) is the composite construction of the residue of keratinocyte closely that separated by the lipid functional areas.Compare with gastric mucosa with the oral cavity, horny layer is more difficult to be made exogenous or endogenous molecule infiltrates through in the body.Horny layer is formed by keratinocyte, and keratinocyte constitutes the major part of epidermis cell, and they lose its nuclear, become horn cell.The cellularity that these are dead horny layer, its thickness as mentioned above, is the very big waterproof membrane of resistance only for about 10-30 μ m, it prevent body be subjected to the invasion and attack of foreign substance and liquid and dissolved molecule to external migration.Coming off and forming new horn cell horny layer is constantly upgraded by when decortication horn cell by the keratinization process.

By changing resistance (diffusion coefficient) or driving force (diffusion gradient), the percutaneous that can increase medicine or analyte flows.Can promote to flow with what is called infiltration or chemical promoter.Chemical promoter is being known in the art, below with for a more detailed description.

Increase skin is electrophoresis to the infiltrative another kind of method of medicine.Electrophoresis relates to that the part that applies external electrical field and ionic drug is carried or carries (electro-osmosis) with the part with the nonionic medicine of the current carrying of ion-transfer.Though promote that with electrophoresis infiltration is effectively, medicine controlled releasing and irreversible skin injury are to follow this technology and the problem that occurs.

Acoustic energy also has been used to promote the infiltration to medicine and other molecule of skin and synthetic film.The ultrasonic mechanical pressure ripple that is defined as frequency above 20kHz, H.Lutz et al., Manual of Ultrasound3-12 (1984).Acoustic energy produces its vibration by piezoquartz or other electricapparatus element in alternating current, R.Brucks et al., 6 Pharm.Res.697 (1089)Use acoustic energy increase skin that the infiltration of drug molecule has been called as sound swimming (sonoporesis or phonophoresis).

Should be able to make molecule from collecting or detect to external by cutaneous metastatic in the body although people know on the penetration theory that promotes skin, still not have feasible method and disclose.The U.S. Patent No. 5,139,023 of authorizing Stanley etc. has disclosed a kind of instrument and method that is used for the non-damage blood sugar test.Use chemosmosis promoter to increase the infiltration of mucosal tissue or skin in this invention to glucose.Then, glucose is through mucous membrane tissue or skin diffusion passively, is received capture medium.Determined and the correspondingly definite blood sugar level of glucose amount in the receiver media.Yet as described in Stanley etc., this method is more effective when being used for mucosal tissue (as BT), makes the glucose that can collect detectable amount at about 10-20 after minute period of delay in receiver media.But the method that Stanley etc. instructed causes extremely long period of delay, according to used promoter compound, is 2-24 hour, then could be at the glucose of vitro detection to the skin through the people (thermal release epidermis) diffusion of detectable amount.May and improve the required time span of stratum corneum barrier penetrating power owing to the passive diffusion of chemosmosis promoter percutaneous Chang period of delay like this, and the passive time span that diffuses out of glucose percutaneous.Therefore, Stanley etc. obviously do not disclose the method for non-damage ground transport through skin blood glucose and other analyte, but these blood electrolyte of this method fast detecting, as for the blood sugar monitoring of diabetics and for a lot of other body analytes required.

It is known using the acoustic energy loading, but the result is disappointed mostly, because permeability improves seldom.Increase the mobile effectiveness of medicine percutaneous for acoustic energy consensus is not arranged as yet.Some research report the success of sound swimming, J.Davick et al., 68 Phys.Ther.1672 (1988); J.Griffin et al., 47 Phys.Ther.594 (1967); J.Griffin ﹠amp; J.Touchstone, 42 Am.J.Phys.Med.77 (1963); J.Griffin et al., 44 Am.J.Phys.Med.20 (1965); D.Levy et al., 83 J.Clin.Invest.2074; D.Bommannan et al., 9 Pharm.Res.559 (1992), other research has obtained the result who negates, H.Benson et al., 69 Phys.Ther.113 (1988); J.McElnay et al., 20 Br.J.Clin.Pharmacol.4221 (1985); H.Pratzel et al., 13 J.Rheumatol.1122 (1986).Adopt the system of rodent skin to show the most promising result, and adopt the general disappointed result of demonstration of system of application on human skin.Those skilled in the art know, and rodent skin is more much bigger than application on human skin permeability, so The above results do not instruct those skilled in the art how to effectively utilize sound swimming, as are applied to through people's skin and carry and/or detect.

The application No.08/152 that awaits the reply that on November 15th, 1993 submitted to; 442 (existing U.S. Patent No.s 5; 458; 140) and the application No.08/152 that awaits the reply that submitted in 8th of December in 1993; 174 (existing U.S. Patent No.s 5; 445,611) disclose and claimed at the detection analyte with utilize the remarkable improvement of acoustic energy on to the body loading, they all draw at this and are reference.In these inventions, the percutaneous sampling of analyte or the percutaneous of medicine are carried by using acoustic energy to combine with chemosmosis promoter and are finished, and the combination of the intensity of used acoustic energy, phase place or frequency or these parameters is modulated.Also disclosed the perforation that causes through acupuncture, water flow jet, laser, electroporation or other method, utilization can have the synthetic acoustic energy of frequency, intensity and/or phase place controllably molecule to be pushed and/or pumps into horny layer.

Promote that the formation of micropore (being micropunch) on the horny layer of drug conveying is the purpose of various researchs, cause the issuing of patent of these technology.

Jacques et al., 88 J.Invest.Dermatol.88-93 (1987) instructs a kind of medication, system adopts wavelength, pulse length, pulse energy, umber of pulse and pulse recurrence rate be enough to the to ablate pulse laser of horny layer and the not obvious damage epidermis below it, the horny layer of ablation skin area, drug administration is in ablated region then.This work causes U.S. Patent No. 4,775, and 361 authorize Jacques etc.By adopting Ultra-Violet Laser irradiation ablation skin more earlier by reports such as Lane, Arch.Dermatol.609-617 (1985).Jacques etc. are confined to use the light and the expensive laser instrument of seldom several wavelength.

The U.S. Patent No. 5 of Tankovich, 165,418 (hereinafter being called " Tankovich ' 418 "), disclosed a kind of method that obtains blood sample, be with one or several laser pulse irradiation human or animal skin, the energy of described laser pulse is enough to cause that the skin histology vaporization extends through the hole of epidermis and cuts at least one blood vessel to produce one on skin, a certain amount of blood is discharged, so that it can be collected through the hole.Therefore, thus Tankovich ' 418 is not suitable for non-damage or damagingly makes horny layer have permeability to make medicine can be transported in the body or make the intravital analyte can be analyzed minimumly.

The U.S. Patent No. 5,423,803 of Tankovich etc. (hereinafter being called " Tankovich ' 803 ") discloses the laser ablation method of the shallow-layer epidermis cell of the application on human skin that is used for cosmetic application.This method comprises uses light absorptive " pollutant " in the epidermis skin, force in these pollutant some to enter cuticular intercellular substance, the skin that soaks into the laser pulses irradiate of sufficient intensity, then the energy of pollutant absorption makes pollutant with enough energy blasts, to tear some epidermis cell.Tankovich ' 803 further instructs, pollutant should have high energy absorption under the wavelength of laser beam, like this, laser beam must be the pulsed light beam that the persistent period is less than 1 μ s, pollutant must be pressed into the epidermis upper strata, pollutant must be with enough energy blasts, to tear epidermis cell after absorbing laser energy.This invention also fails to disclose or point out the method for conveying medicine or collection analyte.

Raven etc. (WO 92/00106) have described the method for removing diseased tissues from the body selectivity, promptly give a kind of chemical compound to selected tissue, the infra-red radiation of this chemical compound high absorption wavelength 750-860nm, should the zone with corresponding illuminated with infrared radiation, its power be enough to cause administered compound tissue the heat vaporization but be not enough to cause the heat vaporization of the tissue of administered compound.Light-absorbing compound should water-soluble or serum, as indocyanine green element, chlorophyll, porphyrin class, contain the chemical compound of haemachrome, or contains the chemical compound of polyene structure, and power level is at 50-1000W/cm ²Scope in or higher.

The heat treatment method of a kind of tumor tissues of Konig etc. (DD 259351) instruction comprises radiating dielectric deposition with absorptive red light and/or near infrared spectrum district on tumor tissues, the tissue that is soaked into the laser irradiation of suitable wavelength.Absorbing medium can comprise methylene blue, porphyrinogen or its condensation product and phthalocyanine blue.This patent has been enumerated methylene blue that strong absorption is arranged at the 600-700nm place and 647 and the Kr laser launched of 676nm place.Power level should be 200mW/cm at least ²

Now prove, use method that adhesive tape was sticked and removed in the same place of skin repeatedly to tear horny layer off from the small size of skin, can easily collect the interstitial fluid of any amount, these interstitial fluids can be used to detect a lot of analyte of interest.Equally, " tear shelled with adhesive tape " skin also is proved to be easily infiltration, can be with in the chemical compound percutaneous input body.Unfortunately, " adhesive tape tears stripping " stays next open wound, and it needs to heal several weeks, so reason and other reason, and not as the acceptable hands-on approach that in using widely, promotes transport through skin.

As discussed above, now prove, pulse laser (as run on 193nm excimer laser, run on the erbium laser of nearly 2.9 μ m or run on the CO of 10.2 μ m ₂Laser instrument) is used in the aperture of ablating out on people's horny layer.These laser ablation technologies are opened by horny layer for selectivity and potential non-damage ground and are carried and/or the method for thief hole provides probability.Yet,, still do not have commercial product at present based on this notion owing to follow the expense of making us hanging back of these light sources.The present invention is by directly importing heat energy cuticular method with very strict space and the temporal resolution that limits, and can adopting at a low price, the energy produces the little ablation of needed horny layer.

In view of the above problems and/or defective, inventing a kind of method that improves percutaneous permeability safely, so that in time frame faster, damage minimum ground or non-damage ground detection body analyte, will be obvious improvement in this area.Another obvious improvement in this area will provide the minimum or non-damage of damage ground and improve the method that medicine enters the percutaneous flow velocity in individual selection zone.

Summary of the invention

An object of the present invention is cuticular barrier to be reduced as far as possible,, make these analytes energy detected so that controlledly collect from intravital analyte in the hole on horny layer with method for punching.

A further object of the present invention provides the method that the micropore on horny layer that combines with acoustic energy, penetration enhancer, barometric gradient etc. detects analyte in the selected body.

Micropore control medicine or other molecule percutaneous that another object of the present invention provides on horny layer flow into the flow velocity that (if desired, flows into blood flow) in the body.

Purpose in addition of the present invention provides the micropore on horny layer that combines with acoustic energy, penetration enhancer, barometric gradient etc. and conducts drugs to intravital method.

These and other objects can be finished by the method that detects analyte concentration in the individuality is provided, and this method comprises by the horny layer in a raising selected zone, surface of the following steps infiltrative step to analyte:

(a) horny layer of institute's favored area is bored a hole, do not cause major injury by on horny layer, forming micropore, thereby reduce the barrier that horny layer is withdrawn from analyte covering weave under it;

(b) collect a selected amount of analyte; With

(c) analyte of collecting is quantitative.

In a preferred embodiment, this method comprises that further the acoustic energy with the frequency in about 5KHz to 100MHz scope puts on institute's favored area of boring a hole, and wherein acoustic energy is modulated in a kind of mode that is selected from frequency modulation, amplitude modulation, phase modulation and combination thereof.In a further preferred embodiment, this method comprises further individual institute favored area is contacted with chemical promoter that application of sonic energy further promotes withdrawing from of analyte.

Cuticular perforation is finished by being selected from following a kind of method: the institute's favored area that (a) span is not more than 1000 μ m areas contacts the horny layer of ablating with a thermal source, selected regional organization bound water and other temperature that can vaporize material are elevated on the evaporation point of water and other material of can vaporizing, thereby remove the horny layer of institute's favored area; (b) use once the micro-knife puncture horny layer of calibrating, to form the micropore of diameter≤about 1000; (c) acoustic energy beam of tight focus is focused on the horny layer horny layer is ablated; (d) with high pressure liquid stream horny layer is carried out the hydraulic pressure puncture with the micropore of formation diameter≤about 1000 with (e) with the micropore of very brief electric pulse puncture horny layer with formation diameter≤about 1000.

The cuticular preferred embodiment of thermal ablation comprises the dyestuff treatment of selected zone that shows the strong effective dose that absorbs at least with paired pulses light emitted scope, and will be on dyestuff from a series of pulse concentrations of light-pulse generator output, dyestuff is heated to is enough to conduct heat to horny layer, and selected regional organization bound water and other temperature that can vaporize material are elevated on the evaporation point of water and other material of can vaporizing.Preferably, light-pulse generator is launched with the wavelength that is not obviously absorbed by skin.For example, light-pulse generator can be in about 630-1550nm scope the emitted laser diode, in about 700-3000nm scope, launch with the optical parametric oscillator of laser diode pumping or be selected from a kind of light source of arc light, electric filament lamp and light emitting diode.Also sensor-based system can be set, in order to determine when cuticular barrier is excluded.Preferable sensor-based system comprises light that reception comes from selected regional reflex and the light collecting device that reflected light is focused on photodiode, the light of collectiong focusing and the photodiode that sends a signal to controller, the catoptrical character of signal indication wherein, and be coupled to photodiode and light-pulse generator with received signal with after the signal of preliminary election receives, cut off the controller of light source.

In a further preferred embodiment, this method further comprises with chiller the cooling of horny layer institute's favored area and proximate skin tissue, is under the steady-state condition of pre-cooling before making described institute's favored area and proximate skin be organized in puncture.

In another preferred embodiment, this method comprises the ablation horny layer, and interstitial fluid is flowed out from micropore, and collection organization's interstitial fluid is also analyzed analyte in the collected interstitial fluid.After collection organization's interstitial fluid, can apply energy from the effective dose of laser diode or other light source, make the interstitial fluid cohesion that is retained in the micropore with closed porosity.Preferably apply vacuum in the collection of institute's favored area of boring a hole with the promotion interstitial fluid.

In also having an embodiment, this method is included in the unfocused rayed institute favored area of using at least before the horny layer perforation from light-pulse generator, uses the favored area sterilization of light-struck institute.

Another preferred embodiment of horny layer perforation comprises institute's favored area is contacted with tinsel, the temperature of institute's favored area is elevated to greater than 100 ℃ from peripheral skin temperature in about 10-50ms, in about 30-50ms, make the temperature of institute's favored area get back to the temperature that is about surrounding skin then, wherein this elevated temperature repeats repeatedly with the circulation of getting back to about peripheral skin temperature, and making is enough to reduce cuticular barrier.Preferably, get back to the step that is about peripheral skin temperature by tinsel being recalled do not contact with horny layer.The device of the electrical impedance of passing through horny layer institute's favored area and proximate skin tissue between monitoring tinsel and the individuality and the device of propelling tinsel position preferably also are provided, make when ablating, when resistance reduces thereupon, propulsion plant contains the propelling tinsel, contacts with horny layer when making the tinsel heating.In addition, preferably provide to make the device of tinsel from recalling with the horny layer position contacting, wherein monitoring device can measure contact the impedance variation of being followed with horny layer following table cortex and transmit signal to withdraw from device with tinsel from recalling with the horny layer position contacting.Tinsel can heat with the Ohmic heating element, current loop with high resistance point can be arranged, wherein use the synthetic electric current that is subjected to that heats by described current loop to modulate the temperature of high resistance point, maybe tinsel can be placed can synthetic magnet exciting coil (excitation coil) AC magnetic field, when the operculum magnetic coil passes to alternating current, can produce the eddy current that is enough to by inner ohmic loss heated wire.

Improving method that active penetrant percutaneous flows into the speed of individual institute favored area comprises by following steps and improves the infiltrative step of the horny layer in a selected zone, surface to active penetrant:

(a) horny layer of institute's favored area is bored a hole, do not cause major injury by on horny layer, forming micropore, thereby reduce the barrier of horny layer active permeate flow to covering weave under it; With

(b) will contact with the compositions that comprises the effective dose penetrant through institute's favored area of perforation, and promote penetrant to flow into body.

In a preferred embodiment, this method further comprises with certain intensity and frequency and applies acoustic energy within a certain period of time in institute's favored area of having bored a hole, thereby this intensity and frequency can be set up the flow of liquid effect effectively and improve the speed that the penetrant percutaneous flows into body.

Also provide a method to be used to apply the institute favored area of tattooing (tatoo), may further comprise the steps to individual surface skin:

(a) horny layer of institute's favored area is bored a hole, do not cause major injury by on horny layer, forming micropore, thereby reduce the barrier of horny layer active permeate flow to covering weave under it; With

(b) will contact with the compositions that comprises effective dose tattooing ink through institute's favored area of perforation, and promote described ink inflow body as penetrant.

Also further provide a method to be used to reduce in skin selection area analyte and be diffused into the time delay of the interstitial fluid of described individuality from the blood of individuality, this method comprises uses chiller in the described selection area of skin.

Also further provide a method to be used to reduce the vaporization and the vapour pressure thereof of interstitial fluid, wherein said interstitial fluid is that the micropore in the cuticular institute favored area of individual's skin is collected, and this method comprises that the application chiller is in the described selection area of skin.

The simple description of several views of accompanying drawing

Fig. 1 is the light of transmission laser diode and the sketch map of monitoring perforation process.

Fig. 2 is the sketch map of the closed loop feedback system of monitoring perforation.

Fig. 3 A is the optics perforation system that comprises chiller.

Fig. 3 B is the top view of illustrating the chiller of Fig. 3 A.

Fig. 4 is the Ohmic heating schematic representation of apparatus with mechanical actuator.

Fig. 5 is a high resistance current loop cross-sectional side view.

Fig. 6 is the schematic representation of apparatus of regulating heating with eddy-current heating.

Fig. 7 is a sketch map of measuring the closed loop impedance detector of degree of perforation with the variation of impedance.

Fig. 8 A-D represents with the CuPc processing, then respectively with power density 4000W/cm ²The cross section of the application on human skin crossed of 0,1,5 and 50 810nm optical pulse irradiation.

Fig. 9-the 11st is at the Temperature Distribution sketch map of simulating with the optics perforation in the hot piercing process.

Figure 12 and 13 is in optics perforation simulation hot piercing process, respectively in horny layer and lived epidermis as the sketch map of the temperature of time function.

Figure 14-16 is shown in respectively with the Temperature Distribution of (being cooled before wherein being organized in perforation) in the optics perforation simulation hot piercing process, temperature in horny layer as the function of time and the temperature in the lived epidermis function as the time.

Figure 17-19 is shown in the Temperature Distribution (wherein organized with the heating of thermometal line) in the simulation hot piercing process, the temperature in horny layer respectively as the function of time and the temperature in the lived epidermis function as the time.

Figure 20-22 is shown in the Temperature Distribution (wherein organizing with the heating of thermometal line and before being organized in perforation and be cooled) in the simulation hot piercing process, the temperature in horny layer respectively as the function of time and the temperature in the lived epidermis function as the time.

Figure 23 and 24 is shown in the Temperature Distribution of (wherein the operating parameter by Tankovich ' 803 carries out the optics heating with tissue) in the simulation hot piercing process and the temperature in the horny layer function as the time respectively.

Figure 25 is interstitial fluid (ISF; O) and blood ( ^*) glucose level is as the diagram of the function of time.

Figure 26 is the distributed points diagram of the ISF glucose of Figure 25 and the difference item between the blood glucose data.

Figure 27 represents that gained ISF is to the rectangular histogram of the relative deviation of blood glucose levels among Figure 25.

Figure 28 represents to carry the cross section of medicine signal conveyer device of institute's favored area on the individual's skin.

Figure 29 A-C is the diagram of delivery of lidocaine to the affected skin area of institute's favored area of horny layer perforation (Figure 29 A-B) or puncherless (Figure 29 C).

Figure 30 represent from single with suction (o) and suction and combination of ultrasound use ( ^*) time interstitial fluid amount collected from micropore comparison curves.

Figure 31,32 and 33 represents ultrasonic vibrator/vacuum equipment, the sectional view of this device and the schematic cross-section of this device that the results interstitial fluid is used respectively.

Figure 34 A-B represents the top view of the ultrasonic vibrator handed and the side view of scraper end thereof respectively.

Describe in detail

Disclose and describe be used to make horny layer to be easy to permeate with this method that promotes the sampling of impart transdermal drug delivery and analyte before, should be understood that the present invention is not limited to specific structure, treatment step and material that this description discloses, because these structures, treatment step and material can some changes.What be appreciated that also has, and the used term of this description is only used for describing the purpose of particular, rather than in order to limit, because scope of the present invention will only be subjected to the restriction of claims and equivalent thereof.

In this description and claims, must be noted that singulative " a ", " an " and " the " comprise plural object, unless context refers else clearly.Therefore, for example mention the method for conveying " medicine ", comprise the mixture of carrying two or more medicine, mention the analyte that " analyte " comprises that one or more are such, mention the mixture that " penetration enhancer " comprises two or more penetration enhancer.

In description of the invention and claim, will be with the following term that meets following definitions.

" perforation " used herein, " micropunch " or any this similar nomenclature be shown on the individual's skin institute favored area horny layer form duck eye or aperture with weaken make analyte under skin surface by for analyze or when making active penetrant or medicine enter body for treatment this layer to their barrier.Preferable is, hole or bore dia are not more than about 1mm, are more preferred from diameter and are not more than about 100 μ m, and extend into horny layer, is enough to destroy the barrier of this layer and the tissue below it is had no adverse effects.

" ablation " used herein the removal of the in check cell that expression is caused by the kinetic energy that discharges when the temperature of vaporization takes place when but the cell vaporized component is heated to, owing to the quick volumetric expansion that this phase transformation produced causes cell and may cause that some adjacent cells " is blown away " from the ablation position.

" puncture " used herein or " micropuncture " expression uses machinery, hydraulic pressure or electric device that horny layer is bored a hole.

Finish in " ablation " or " puncture " in the scope of same perforation purpose (promptly form hole or hole on horny layer and below the not obvious damage tissue), these terms are used interchangeably.

" penetrating (penetration) promotes " used herein or " infiltration (permeation) promotes " expression skin increase the permeability of medicine, analyte, dyestuff, coloring agent or other chemical molecular (being also referred to as penetrant), permeate cuticular speed thereby increase medicine, analyte or chemical molecular, with promote cuticular perforation, analyte withdraw from by horny layer or medicine through horny layer carry and enter below tissue.Can observe by the promotion osmosis of using such promoter to produce, as pass through the diffusion of animal or human's skin as the dyestuff of penetrant with the diffusion observation of use instrument.

" chemical promoter " used herein, " penetration enhancers ", " penetration enhancer " etc. comprise can increase penetrant, analyte or other molecule through mobile all promoter of skin, is only limited with function.In other words, comprise chemical compound and solvent and other chemical promoter that all make the cell membrane disorder.

" dyestuff " used herein, " coloring agent " etc. use interchangeably, refer to show the strong biologically suitable chromogen that absorbs in the transmitting boundary of cuticular tissue with the light-pulse generator that forms micropore that be used for ablating.

" percutaneous " used herein or " percutaneous " expression penetrant penetrates skin and passes through skin, reach medicine and effectively treat blood concentration or deep tissue level, or the molecule that exists in the body (" analyte ") comes out by skin, so that can be at external collection analyte molecule.

Term used herein " penetrant ", " medicine " or " pharmacologically active agent " or any other similar term are represented to be suitable for the method known to before this area and/or any chemistry or the biomaterial or the chemical compound of the method transdermal administration that the present invention instructed, it causes needed physiology or pharmacodynamics effect, it can include but not limited to that (1) has the prophylactic effect to body, prevent undesired physiological effect such as prevention infection, (2) symptom that palliates a disease and cause, as alleviating pain or the inflammation that produces as the result of disease, and/or (3) are alleviated, alleviate or eliminate fully the disease of body.This effect can be partial, as the local anesthesia effect is provided, and maybe can be whole body.The present invention does not relate to the new penetrant or the activating agent of newtype, and is limited to existing activating agent in this area or penetrant or can be confirmed as activating agent later on and the mode of movement of the material that is suitable for carrying with the inventive method.Such material comprises various types of chemical compounds of (comprising by body surface and film (comprising skin)) in the common input body.In general, include but not limited to: anti-infective, as antibiotic and antiviral agent; Analgesic and analgesia mixture; Anorexigenic; Anthelmintic; Anti-arthritic; Antasthmatic; Anticonvulsant; Antidepressants; Antidiabetic drug; Diarrhea; Antihistaminic; Antiinflammatory; The migraine preparation; Antinauseant; Antineoplastic agent; Drug for parkinson's disease; Antipruritic; Psychosis; Antipyretic; Spasmolytic; Anticholinergic agent; Sympatheticomimetic; Yellow fast quinoline derivant; Cardiovascular preparation comprises potassium and calcium channel blocker, beta blocker, alpha block agent and anti-arrhythmic; Antihypertensive; Diuretic and antidiuretic; Vasodilation comprises whole body blood vessel, coronary vasodilator, peripheral blood vessel and cerebral vasodilator; Central nervous system's stimulant; Vasoconstrictor; Cough and cold-treating preparation comprise that releasing is towards the blood agent; Hormone as estradiol and other steroid, comprises corticosteroid; Hypnotic; Immunosuppressant; Muscle relaxant; Anticholinergic agent; Psychostimulant; Tranquilizer and tranquilizer.Use method of the present invention, no matter ionizing and nonionic chemical medicine thing, no matter high molecular or low-molecular-weight drug all can be carried.

The pharmacologically active agent of " effectively " used herein amount refers to that the chemical compound of q.s provides needed part or general action with rational interests/danger than (this is that any medical treatment has).The infiltration or the chemical promoter of " effectively " used herein amount refer to a selected amount, with increase and needed length of penetration, the speed of administration and the amount of the medicine of being carried that needed percutaneous permeability is provided.

" carrier " used herein or " excipient " refer to be fit to use with the other medicines active substance, the carrier mass of no obvious pharmacologically active under employed quantity, comprise any this material known in the art, as any liquid, gel, solvent, liquid diluent, solubilizing agent etc., it is nontoxic in applied amount, can not interact with deleterious mode and the medicine of preparing to give.The example of suitable carriers used herein comprises water, Dormant oils, siloxanes, inorganic gel, aqueous emulsion, liquid sugar, wax, vaseline and various oil and polymeric material.

" biomembrane " used herein is meant the membrane material that exists in the body of living, it is separated a zone and another zone of body, under many circumstances body is separated with its environment facies.Therefore, comprise skin and mucosa.

" individuality " used herein refers to it is implemented humans and animals of the present invention.

" analyte " used herein refers to be suitable for the technology of instructing with the present invention or technology known in the art by biomembranous any chemistry or biomaterial or chemical compound, and body may be wondered its bulk concentration or activity one by one.Glucose is the object lesson of analyte, because it is the analyte that is adapted to pass through skin, individual (for example those suffer from the people of diabetes) may wonder their glucose blood concentration.Other example of analyte includes, but is not limited to such as chemical compounds such as sodium, potassium, bilirubin, carbamide, ammonia, calcium, lead, ferrum, lithium, Salicylates.

" percutaneous flow velocity " used herein is the skin effusive speed of any analyte by individual (human or animal), or any medicine, pharmacologically active agent, dyestuff or pigment enter and pass through the speed of the skin of individual (human or animal).

Term used herein " intensity amplitude ", " intensity " and " amplitude " are used with the free burial ground for the destitute, refer to the amount with the energy of acoustic energy system generation.

" frequency modulation " used herein or " frequency sweep " refer to amplitude or frequency variation continuously, step by step or progressively in cycle regular hour.Frequency modulation be in the cycle regular hour frequency step by step or the variation of step, as in 1 second, from 5.4 to 5.76MHz, or in 0.1 second 5-10MHz, or in 0.1 second 10-5MHz, or be suitable for concrete any other frequency range or the time cycle of using.Complex modulated can comprise that frequency and intensity change simultaneously.For example, U.S. Patent No. 5,458,140 Fig. 4 A and 4B can represent respectively simultaneously single acoustic energy oscillator is applied amplitude and warbled situation.

" phase modulation " used herein expression is as U.S. Patent No. 5,458,140 shown in Figure 4, the timing of signal concerns corresponding to its starting stage and is changed.The frequency of signal and amplitude can remain unchanged.With variable delay, as make signal corresponding before it state or postpone or shift to an earlier date corresponding to another signal-selectivity ground is temporary transient, can finish phase modulation.

As described herein, acoustic energy is in its various application, and as frequency modulation, amplitude modulation or phase modulation or its combination, and chemical promoter is used in combination with synthetic acoustic energy, can change in the frequency range between about 5kHz to 100MHz, be good with the scope between about 20kHz and the 30MHz.

" non-damage " used herein expression need not to insert body with pin, intubate or other damaging medical apparatus.

" damaging minimum " used herein refers to use machinery, hydraulic pressure or electric installation damage horny layer not to cause to produce duck eye or micropore the substantial damage of organizing below it.

The method that is used for the horny layer perforation

The existing the whole bag of tricks in the available this area of the formation of micropore and their some modification methods of herein disclosing are finished in the horny layer.

It is a kind of with the excimer laser cuticular method of ablating that the use of the laser ablation that (the same) such as Jacques etc. (in U.S. Patent No. 4,775,361 in) and Lane described provides undoubtedly.Found that at wavelength 193nm and pulse width 14ns, available each laser pulse is with about 70-480mJ/cm ²Irradiation will about 0.24-2.8 μ m horny layer remove.Along with the increase of pulse energy, more tissue is removed from horny layer, bores a hole required pulse fully then still less on this layer.In the thermal relaxation time limit, must be absorbed to produce suitable micro-explosion and to cause the lower end threshold value of the irradiation of tissue ablation in 50 milliseconds (ms), to be about 70mJ/cm by horny layer ²In other words, must in 50 milliseconds of windows, transmit total 70mJ/cm ²This can use 70mJ/cm in 50 milliseconds time ²Individual pulse or 10 7mJ/cm ²Pulse or 1.4W/cm ²Continuous irradiation finish.The upper limit of irradiation is such amount, and it does not damage following tissue with the ablation horny layer, can rule of thumb be determined by light source, optical wavelength and other variablees in those skilled in the art's experience and knowledge scope.

" transmission " speech is meant that the tissue that the energy of described quantity is ablated by need absorbs.In the situation of excimer laser wavelength 193nm, cuticular tissue has partially absorbed 100% energy basically for uppermost 1 or 2 micron.Suppose about 20 micron thickness of horny layer, at longer wavelength, for example at 670nm, only about 5% incident illumination is absorbed in 20 micron layers.This means that about 95% high power-beam has entered the tissue below the horny layer, there, it might produce obvious damage.

Ideal method only is to use the power for necessary amounts to come horny layer bored a hole can not cause following organize hemorrhage, heating or other damages, from following tissue with extraction of analytes or transmission medicine or other penetrants.

It will be more favourable using than using from the more economical energy of the energy of excimer laser, thereby also more useful.The required expense of the excimer laser of the light of running and maintenance emission far ultraviolet zone wavelength is big more than the diode laser of the light of for example visible emitting and region of ultra-red wavelength (600-1800nm).Yet at long wavelength, horny layer little by little becomes more transparent and absorbs and mainly occurs in the following tissue.

The invention provides the method for removing cuticular barrier function fast and painlessly, to promote when the local dispenser therapeutant percutaneous to be delivered in the body or to choose in vivo for the analyte of analyzing.This method is used the program that contacts beginning with the small size thermal source with cuticular target scope.

Thermal source must have the several important performances that below will discuss.At first, the size of thermal source must be suitable, and general diameter is about the 1-1000 micron, so that with in contacting of skin is limited among a small circle.Second, thermal source must be in cycle time be adjusted to the cuticular temperature of contact point more than 123 ℃ from surrounding skin surface temperature level (33 ℃), falls after rise then to reducing to minimum near the surrounding skin surface temperature will and make feeling that the subject produces to the damage of activated tissue.This adjusting available electron, machinery or chemical method are carried out.

In addition, if thermal source has enough little thermal mass and limited energy source to promote its temperature, thereby when its with water content when the tissue more than 30% contacts, heat in these tissues is scattered to be enough to the maximum temperature of thermal source is limited in below 100 ℃, then helps bringing into play the inherent degree of depth limited features of micropunch method.In case thermal probe passes the lower floor that horny layer enters epidermis, this feature can stop the heat vaporization effectively.

With regard to regard to the thermal source of contact skin, it is by a series of one or more thermoregulations circulations, from the peak temperature more than the initial peripheral skin temperature to 123 ℃ again near peripheral skin temperature.For make micropunch method subject feel reduce to minimum or make its elimination, the persistent period of these pulses is restricted, the dermal tissue cooling that is enough to make activated organized layer in the skin especially weak is grown at interpulse interval, so that mean temperature is about below 45 ℃.These parameters according to thermal probe and below the thermal time constant that active epidermal tissue is arranged (about 30-80 millisecond) between the weak tissue and deciding in the corium.The above-mentioned result who uses pulse heat energy is, the horny layer in the very little target spot of enough energy input is arranged, and like this, the local temperature of this portion of tissue is promoted to the vapourizing temperature that enough is higher than the moisture that is organized constraint in the horny layer.When temperature rises to more than 100 ℃, vaporization will take place and very rapidly expand in the moisture of this localization point endocuticle (being generally 5-15%), cause that the horn cell near the horny layer of this vaporization zone is removed by steam.U.S. Patent No. 4,775,361 point out, 123 ℃ horny layer temperature is represented the threshold value that the type flash distillation takes place.When one after the other using heat pulse, be that horny layer further was removed before clear layer was formed with micropore at layer from horny layer to the below the epidermis.Be limited in below the thermal time constant of epidermis by the persistent period with thermal pulse and make the heat energy that enters epidermis have the sufficiently long time to dissipate, the intensification of activated layer is atomic in the epidermis.This makes whole micropunch process not have any sensation and following and tissue is not had under the situation of any damage and carry out the subject on every side.

The present invention includes a kind of method of making the micropore of wide about 1-1000 micron at the horny layer of fell painlessly.The key of successfully implementing this method is to make the appropriate thermal energy instant heating probe that contacts with horny layer.The primary technology that requires in the manufacturing of suitable thermal probe is the such device of design, and it can keep can sufficiently high frequency carrying out thermal conditioning with required the contact also of skin.

Can make suitable thermal probe by the suitable light-absorbing compound of horny layer local application such as dyestuff or coloring agent are waited, selected chemical compound should be able to absorb the light of the wavelength of selected light emitted.In the case, the selected light source wavelength that can be its emission the laser diode that generally can not absorbed by skin histology.By light-resource fousing is also changed the intensity that focuses on the luminous flux on this on the point of dyestuff local surfaces, can carry out temperature modulation to target region.Toward horny layer local application suitable light-absorbing compound such as dyestuff or coloring agent etc. (selected chemical compound should be able to absorb the light of the wavelength of lasing light emitter emission), can use the energy of its emission wavelength by earlier than the longer lasing light emitter of excimer laser.Same notion is applicable to any wavelength, and what the technical staff will do only is to select suitable dyestuff or coloring agent and optical wavelength.The technical staff only need consult any reference manual can find the suitable dyestuff and the maximum absorption wavelength of this dyestuff.Such list of references has the The Sigma-AldrichHandbook of Stains of Green, Dyes and Indicators, Aldrich Chemical Company, Inc.Milwaukee, Wisconsin (1991).For example, CuPc (dyestuff indigo plant 15; CPC) absworption peak is near 800nm; The absworption peak of CuPc tetrasulfonic acid (acid blue 249) is near 610nm; The absworption peak of indocyanine is near 775nm; The absworption peak of koha is near 703nm.CPC is especially suitable to present embodiment, and it be the reasons are as follows: it is very stable and inert chemical compound, is allowed to be used as dyestuff in transplantable suture by U.S. food Drug Administration; It is very strong in the absorption of wavelength 750-950nm, is fit to many emitters of solid cheaply, and as laser diode and LED, in addition, the wide zone of its light belt can directly not absorbed by skin histology significantly yet; The vapourizing temperature of CPC very high (in a vacuum greater than 550 ℃) directly is transferred to gas phase and without liquid phase from solid phase; The thermal-diffusion constant of CPC is less, order focus on thereon luminous energy only optionally heating just in time in the zone of focus and seldom by " focus " peripherad CPC diffusion, thereby help the space orientation of contact thermal probe.

The purpose of this announcement does not really want to list the suitable dyestuff or the detailed bill of coloring agent, because those skilled in the art can easily be determined according to the data that can obtain easily.

This also is same correct for any required concrete light-pulse generator.For example, this method can machinery electric filament lamp that open and close, that focus on as light-pulse generator.Extensive stock catalogue and sales data shown many can be at the laser instrument of near ultraviolet, visible light and near infrared region work.Representational laser instrument is a PLP-02 type Hammamatsu photonic system, and it is 2 * 10 at the output of wavelength 415nm ^-8J; PLP-05 type Hammamatsu photonic system, it is 15J at the output of wavelength 685nm; The SDL-3250 switched pulse train laser of SDL company, near its output wavelength 800-810nm is 2 * 10 ⁶J; The SDL-8630 type of SDL company, near its output wavelength 670nm is 500mW; The single-phase laser instrument of AR-081-15000 type, it is 15 at the output of wavelength 790-830nm, 000mW; Toshiba AmericaElectronic TOLD9150 type, it is 30mW at the output of wavelength 690nm; The Diolite800-50 type of LiCONIX company, it is 50mW at the output of wavelength 780nm.

In the present invention, the pulse laser light source can be at about 100nm to 12, the wide wave-length coverage divergent-ray of 000nm.The transmitting boundary of excimer laser is between about 100 to 400nm.The wave-length coverage of the commercial excimer laser of selling is between about 193 to 350nm at present.The transmitting boundary of laser diode is preferably between about 380 to 1550nm.Through the transmitting boundary of the laser diode of frequency multiplication between about 190 to 775nm.Use the optical parametric agitator of laser diode pumping, then can utilize the longer wavelength region may between about 1300 to 3000nm.Expection is along with laser technology research carries out in a deep going way, and these zones will further be expanded from now on.

Energy transmission or that absorb might not need to obtain from laser instrument, because any light source all can use, no matter be from short-arc lamps such as laser instrument, xenon flash lamp, electric filament lamp, light emitting diode (LED), the sun, or any other light source.Therefore, the concrete instrument that is used for transmitting electromagnetic radiation is important not as relative wavelength and energy.Any suitable apparatus that can transmit institute's energy requirement at suitable wavelength (promptly at about 100-12, between the 000nm) all can be thought to belong within the scope of the present invention.Its essential feature is that energy must be absorbed by light-absorbing compound so that its localization ground heats, and sufficient heat is conducted to the tissue of ablation then in the time frame that allows.

In an illustrative embodiment, thermal probe self is formed by following thin layer: its preferably about 5-1000 micron thickness, form by solid abiotic reactive compound, and be applied in the selection area of subject's skin, this selection area is sufficiently big, can cover the position of waiting to make micropore.The selection of the concrete prescription of chemical compound requires: it has very strong absorption in choosing for light-absorbing compound provides the spectral region of the light source of energy.Probe can be the small pieces of for example solid chemical compound, the film of using the high-melting-point absorption compound to handle, perhaps light-absorbing compound is applied directly on the skin as precipitate or as the float in carrier.Regardless of the structure of light absorption thermal probe, its lateral heat diffusion coefficient must be sufficiently low, and any local heating of make suring remains in the restricted portion, and the main mode of thermal losses will be directly to conduct by the contact point between skin and the probe to enter horny layer.

Popping one's head in required thermoregulation can be by realizing the intensity of light-resource fousing at light-absorbing compound and this light source of adjusting.If the systemic energy of irradiation area is sufficiently high, it will cause that light-absorbing compound promptly generates heat.The energy of transmission and light-absorbing compound all can easily be regulated by the pulse width and the peak power that change light source at the rate of heat addition and the peak temperature of focus.In the present embodiment, have only light-absorbing compound that fraction is focused, the incident optical energy heating to constitute thermal probe, the light-absorbing compound that is applied in than bigger zone, practical puncture position in addition is incidental.By using higher solid phase light-absorbing compound of fusing point such as CuPc (it still is being in solid phase up to the temperature more than 550 ℃) etc., the temperature of thermal probe can promptly rise to hundreds of degree centigrade, and still keeps and contact skin, makes this heat energy conduct and enters horny layer.In addition, present embodiment comprises the light source that selection is such: in its emission spectra, energy generally seldom is absorbed in skin histology.

In case the light-absorbing compound of local application on it arranged in target region, then be activated when light source, and the beams focusing waist is when overlapping with the region surface of handling, thermal probe just can form.The amount of the absorption that takes place in energy density that focuses on waist and light-absorbing compound is set at and is enough to make in several milliseconds the temperature of the light-absorbing compound in the point zone of being determined by the light source focus to rise to more than 123 ℃.When the thermal probe temperature rose, conduction entered cuticular power transfer and organizes to these, and cuticular local temperature is risen.When enough power transfer to this among a small circle horny layer and cause local temperature to be promoted to the boiling point of water contained in these tissues when above, above-mentioned waterishlogging is given birth to flash distillation, ablates at the horny layer of this point.

By opening or closing light source, the selectivity that can promptly regulate the temperature of thermal probe and reach these tissues is ablated, thereby can make big or small point-device hole, and these holes are the uppermost 10-30 micron of transdermal only optionally.

Another feature of present embodiment is, by selecting the general light source that is seldom absorbed by skin or following tissue of its energy and be designed to have the aperture of sufficiently high numerical value by transmission optics system that focus optical is unified, then a small amount of transmission light that is absorbed by thermal probe self does not go deep into depths in the body along with it and disperses.Since the transmission wavelength absorption is seldom arranged, therefore, essentially no energy directly from light sources transmit to skin.Coupling can be in tissue this three-dimensional dilution be since absorb in the beam divergence and the tissue that is untreated low-level due to, cause light beam and organize between complete benign interaction and can not produce any damage.

In a preferred embodiment of the present invention, laser diode is used as the light source that emission wavelength is 800 ± 30nm.Can bring the formation thermal probe by the transparent adhesive tape that local application had been handled, binding agent one side of this adhesive tape has the big speckle of 0.5cm that is formed by CuPc (CPC) fine powder deposition.The absorptance of CPC in the 800nm spectral region is very big, absorbs more than 95% of laser diode radiation emitted energy usually.

Fig. 1 has shown that the optical transmission that is used for this type of laser diode is launched is to the selection area of subject's skin and the system 10 that is used to monitor the perforation procedure progress.This system comprise the intensity of controlling light pulse, persistent period and controller 18 at interval and with its laser diode 14 of closing of Rhizoma Nelumbinis mutually.The light beam 22 of collecting lens 26 is pointed in the laser diode emission, and collecting lens 26 focuses on light beam on the mirror 30.Light beam is reflexed on the object lens 34 by mirror 30 then, and object lens 34 focus on light beam in the pre-reconnaissance 38.This pre-reconnaissance and xyz object stage and connect the planes overlapping in thing hole 46 like this, can shine the selection area of subject's skin.The xyz object stage is connected with controller, like this, and the position of may command xyz object stage.This system also comprises monitoring system, and described monitoring system comprises and the monitor 54 Rhizoma Nelumbinis CCD camera of closing mutually.The confocal calibration of CCD camera and object lens, like this, the progress of perforation procedure can monitoring with the naked eye on monitor.

In another illustrated embodiment of the present invention, be provided with system with the photodiode and the condenser of the confocal calibration of ablative light sources.Fig. 2 shows the sensing system 60 that is used for this embodiment.This system comprises the light source 64 of launching light beam 68, and light beam 68 focuses in the pre-reconnaissance 76 (for example subject's skin surface 80) by optical transmission system 72.Be reflected with a part of light of contact skin, other light are from the irradiation area scattering.The part of the light of this reflection and scattering then focuses on the photodiode 92 by condenser system 88 by filter lens 84.Controller 96 Rhizoma Nelumbinis are closed in laser diode and photodiode, control the output of laser diode respectively and survey the light that arrives photodiode.Only there is selected part to pass through light filter from the spectrum of skin scattering.By analyzing from the skew of the light of target region reflection and scattering, whether system is detectable goes out horny layer and breaks, feeds back with this then and controls light source, cancels light pulse when cuticular micropunch is finished.By using the active closed loop feedback system of the type, can obtain device self regulation, can be general, it produces uniform micropore in horny layer, no matter and whether the subject there are differences, in addition, its power requirement is minimum.

In another illustrative embodiment, added chiller at system to the interface of skin.Fig. 3 A is its sketch map.In this system 100, light source 104 (Rhizoma Nelumbinis is closed in controller 106) emission light beam 108, light beam is by optical transmission system 112 line focusing of going forward side by side.Light beam is focused in the pre-reconnaissance 116 (for example selection area 120 of subject's skin) by optical transmission system.Chiller 124 (for example Peltier refrigerator or other coolers) contact skin makes the skin surface cooling.In a preferable example (Fig. 3 B) of chiller 124, a centre bore 128 is arranged, the light beam of focusing passes this hole contact skin.Referring to Fig. 3 A, heat abstractor 132 is also contacted with chiller again.By its foraminate chiller in center that overlaps with the focus of light is provided, need the interior skin device of gamut of perforation can be precooled to 5-10 ℃.This is pre-cooled to make system that operation has bigger margin of safety, because compare with non-refrigerative embodiment, it significantly reduces to user's cryptesthesia with to the probability of any indirect injury of the epidermis under the site of perforation.And with regard to monitoring, it is minimum that pre-cooled vaporization with interstitial fluid reduces to, and also can provide useful physical property, and for example the surface tension of these interstitial fluids reduces.Also have, known chiller can cause the local increase of the blood flow of these tissues that are cooled, thereby promotes that analyte spreads to interstitial fluid from blood.

This method wherein, is applied to the zone that needs ablation with light-absorbing compound/thermal probe also applicable to other little surgical technics, optionally regulates the probe temperature of selected target site then with light source, acts on tissue by the vaporization-ablation process that produces.

Another feature of the present invention is to help sealing in the ineffective back of micropore with light source.Specifically, under the situation of monitoring internal analysis thing, make micropore and extract some interstitial fluids by this hole.After collecting the interstitial fluid of q.s, under the condition that reduces power, restart light source, to help interstitial fluid rapid condensation in micropore.By liquid is condensed in the hole, this hole in vivo is closed, thereby has reduced infected danger.In addition, all using light source when forming micropore and it is sealed is a kind of method of automatic sterilizing, need not any device is actual penetrates in the body.Also have, the heat stress that is brought out by luminous energy can kill any microorganism that may be present in the ablation position.

The germ-resistant notion of this optics can be extended in this process and to add a step in addition, promptly earlier uses light source in unfocused mode, covers target region, and its irradiation area is bigger 100 microns or more than the micropore effective aperture that will produce.By selecting to use the zone of unfocused light beam in the above, pharosage can correspondingly be reduced to be lower than ablation threshold but to be enough to carries out germ-resistant proper level to skin surface effectively.Make big zone have that the exposure long enough then becomes system configuration the Investigation of Ablation Mode of clear focusing after the time under the light beam of bactericidal action with a kind of successive step or with a series of pulses, optics micropunch process begins at this point.

Another illustrated embodiment of the present invention is to form required thermal probe by metal solid (for example tinsel of minor diameter).Embodiment is such as described above, the contact surface of thermal probe must be adjusted to the temperature more than 123 ℃ from peripheral skin temperature (33 ℃) in the required time, at high temperature the time of Yun Xuing (turn-on time) preferable between about 1 to 50 millisecond, the time of Yun Xuing (turn-off time) is at least about 10 to 50 milliseconds at low temperatures.Especially, if can in the turn-off time of " connection " time about 5 milliseconds and 50 milliseconds with more than the thermoregulation to 150 ℃, then can produce very effective thermal ablation and the subject has only seldom or numbness.

Can successfully implement several adjusting tinsel probe contact area method of temperature.For example, available outer thermal element (for example Ohmic heating element that uses in the soldering iron tip) tinsel that length is short is heated to required high temperature.Fig. 4 is the Ohmic heating device 140 of band mechanical actuator.The Ohmic heating device comprise with tinsel thermal probe 148 mutually Rhizoma Nelumbinis close ohm thermal source 144.Ohm thermal source also pass insulating fixing piece 152 and mechanic adjustment unit 156 (for example solenoid) mutually Rhizoma Nelumbinis close.In this structure, can reach limit, wherein the tip of tinsel probe will be stabilized in by the i.e. a certain equilibrium temperature determined of temperature, tinsel length and diameter, tinsel ambient air temperature and the composition material wiry of ohm thermal source of structural physical parameter.In case reach required temperature, the thermoregulation of the selection area of subject's skin 160 promptly directly carries out with following method by mechanic adjustment unit: alternately with tinsel thermal spike head and 5 milliseconds of turn-on times of contact skin (best), then it is removed, in air, place (best) 50 milliseconds of turn-off times.

Another illustrative embodiment (Fig. 5) is to comprise and controller 178 device 170 of the power supply 174 that closes of Rhizoma Nelumbinis mutually.Power supply with comprise that the current loop 182 of tinsel 186 links, tinsel 186 forms the structure with high resistance point.Be preferably, tinsel is fixed on the fixture 190, and insulator 194 separates the different piece of current loop.Can carry out required thermoregulation as long as regulate by electric current wiry then.It is enough to obtain thermolysis (being connected with power supply by electrode) suitable and that provided by electrode as if the size with tinsel element thermal mass, and then the time of warming of tinsel element and the time of turning cold can be controlled in several milliseconds.Tinsel heats horny layer with the meeting that contacts of the selection area of skin 198, thereby reaches selected ablation.

In Fig. 6, shown with the another illustrative embodiment of hot wire with the horny layer perforation.In this system 200, tinsel 204 can be placed in the adjustable alternating magnetic field that is formed by magnet exciting coil 208.To the logical alternating current of going up of magnet exciting coil, can in the tinsel thermal probe, induce the vortex flow of sufficient intensity, and thermal probe will directly generate heat by the controller 212 that links with magnet exciting coil by inner ohmic loss.This is the scaled-down version that is usually used in tool heads is heat-treated or counter electrode carries out the heating system of degassing processing in vacuum tube or flash tube basically.The advantage of induction heating is to carry out strict control and can easily regulate by the Electronic Control to magnet exciting coil transferring to energy in the tinsel thermal probe.If the thermal mass of tinsel probe self and the cuticular thermal mass that contacts with probe tip are known, then by can very accurately control the temperature of the point 216 that contacts with skin 220 to the control of the induction energy that transmits.Because skin histology is non magnetic at the lower frequency that can carry out eddy-current heating basically, if use the frequency of selecting through suitably in magnet exciting coil, then this alternating electromagnetic field will have no effect to skin histology.

If adopt the contact of machinery control to regulate, the simple closed-loop control system by the impedance that adds between a probe tip and the subject's skin is monitored can realize another feature of the present invention.In this mode, can make probe and subject's contact skin, the rank formula of getting over by resistance reduces to show and comes in contact, and then probe remained on the skin to required " turn-on time ", probe can be removed then.The linear actuator of several types is suitable for the closed loop control of this form, for example voice coil loudspeaker voice coil mechanism, simple solenoid, have the rotary system of cam or bell crank etc.Its advantage is the carrying out along with thermal ablation, and thermal probe also can correspondingly pierce skin, guarantees all the time to contact with the good of skin, thereby realizes effective transmission of required heat energy.In addition, the variation of horny layer and epidermis conductivity provides by closed loop confirms the excellent way whether perforation procedure is finished, and, when the resistance demonstration has arrived epidermis, can stop perforation that is.

Fig. 7 is the illustrative embodiment of such closed loop impedance monitor.In this system 230, ohm thermal source 234 that links with tinsel thermal probe 238 is arranged.This thermal source is fixed on the mechanically-operated controller 246 by insulating fixture 242.Controller 250 links with tinsel and skin 254, the impedance variation of its middle controller detection of skin selection area 258, and when reaching predeterminated level, controller stops perforation procedure.

Miniature lancet and hydraulic piercing device belong to same design, and this lancet is adjusted to and only penetrates horny layer to grant penetrant (for example as medicine) by the hole that forms or to be used for analyzing by hole extraction analyte.Compare with non-damage type device and/or technology, such device is considered to " damage minimal type ".It is well-known using miniature lancet to thrust and extracting blood below the horny layer.These devices can be buied from manufacturer (for example Becton-Dickinson company and Lifescan company) there, and can be used by the control paracentesis depth in the present invention.As the example of the miniature lancet that is used to collect body fluid, can be referring to the PCT application WO 95/10223 (date of publication: April 20 nineteen ninety-five) of Erickson etc.This application shown a kind of be used to puncture to dermal layer of the skin to collect the device that body fluid blood supply sugar concentration etc. is checked, this device can not thrust subcutaneous tissue.

The horny layer perforation also can realize by using ultrasonic unit.Ultrasonic perforating is the change of above-mentioned Optical devices, and different is transfers to the cuticular ablated area that needs with the narrow beam of the very dense of ultrasonic energy, to replace using light source.Required energy is same level, and it is 70mJ/cm that threshold value promptly still must be arranged ²The energy of/50ms is absorbed.Can be with (combination of the intensity of this ultrasonic energy, phase place or frequency or these parameters be conditioned at the transmission ultrasonic energy, extract analyte sample or percutaneous dosing with percutaneous) middle use the same, use and the U.S. Patent No. 08/152 of applying for as mother, the identical pulse concentration ultrasonic transducer of describing in 442 and 08/152,174 transmits the required energy density of ablating.This has the following advantages, can be used for promoting medicine, make it see through horny layer, or body fluid be attracted to skin surface for the identical transducer of analyzing, be used at first making micropore.

In addition, can carry out electroporation and maybe the short pulse of current group with enough energy can be transferred to horny layer, form micropore.Electroporation can be used to manufacturing hole in biomembrane, thereby is known in this area, and electroporation apparatus is on sale on market.Therefore, those skilled in the art can select used instrument and condition to need not to carry out too much experiment according to the policy that provides herein.

The micropore of making in horny layer with method of the present invention makes macromolecule treatment chemical compound transmit by the high flow rate percutaneous.In addition, these lead to intravital atraumatic micropore mouth can touch intravital various analyte, thereby can be used to analyze definite their concentration in vivo.

Embodiment 1

In the present embodiment, prepare the skin sample as follows.Thermal release method (88 with Klingman and Christopher Arch.Dermatol.702 (1963)) isolate the epidermis film from people's corpse whole bark.Described thermal release method comprises whole skin 60 seconds of exposure under 60 ℃ temperature, then horny layer and part epidermis (epidermis film) is peeled off from corium lightly.

Embodiment 2

The thermal release horny layer sample that will make by the method for embodiment 1 is cut into 1cm ²Section.Then these small samples are placed on and make it cling on the cover glass, use the disk that the back side scribbles contact adhesive again on the skin sample, there is the hole of a 6mm at the disk center, facing to tuerculoderma.Sample promptly can be used for experimental test.In some cases, make the skin sample in neutral buffering phosphate solution or pure water, soak a few hours and make its hydration.

As test, emission wavelength is applied on these samples with the output of several different infra-red laser diodes of 1550nm about 810,905,1480 these untreated skin samples.It is 0.4 final object lens that optical transmission apparatus is designed to numerical aperture, produces 25 microns wide waists, records general power that the 905nm that can continuous wave (CW) mode operates and 1550nm laser diode transfer to focus between 50 to 200 milliwatts.905nm and 1550nm laser diode are designed to and can produce the long high-peak power pulse of about 10-200 nanosecond up to the repetition rate of 5000Hz.The peak power level that records pulse laser is 45 watts at 905nm, is 3.5 watts at 1550nm.

Under these operating conditions, any laser does not all have positive effect to the skin sample.60 seconds of Continuous irradiation target region, check at microscopically then, do not find observable effect yet.In addition, sample is placed in the improved Fu Langzi box (Franz cell) (it is commonly used to test with chemosmosis promoter is the percutaneous transmission system of base), before shining and afterwards, a side to the conduction of opposite side from film is measured, found no any variation with laser.According to these tests that the skin sample that obtains from 4 different donors is carried out, can draw to draw a conclusion: at these wavelength, Rhizoma Nelumbinis is closed the luminous energy that enters skin histology to be so little so that not to detect any effect.

Embodiment 3

For assessing subject alive cryptesthesia during by optical energy irradiation under the condition of embodiment 2, the output of each lasing light emitter is applied on finger tip, forearm and the back of the hand of 6 volunteers.When being 810,905 and during 1550nm laser, the imperceptible laser of subject is on-off when.When being 1480nm laser, in the process of shining with 70 milliwatt continuous waves with 1480nm laser, the subject has some sensation, after the short time, forms very little blister because the 1480nm ray is absorbed by one in the water absorption band below skin.Obviously, absorbed energy is enough to bring out blister and forms, but is not enough to the horny layer ablation is removed.In addition, the absorption of 1480nm light mainly appears in (85-90% water content) tissue darker, hydration fully of epidermis and corium, but not in cuticular (10-15% water content) tissue of doing.

Embodiment 4

Confirmed the skin of native state is lacked effect (embodiment 3) afterwards, the energy that a series of chemical compounds absorption luminous energy will absorb has then been assessed by the effectiveness that conduction is transferred in the cuticular target tissue.The chemical compound of test comprises prepared Chinese ink; Black, blueness and red symbol pen that the wiping of " SHARPIE " board is not fallen; Methylene blue; Pinkish red; Epolite #67 (a kind of be used for being molded into the light-absorbing compound that polycarbonate lens is used as laser light protective goggles); Iodine tincture; Iodo-polypyrrole alkane ketone complex (" BETADINE "); CuPc; With the seal China ink.

Use 2 kinds of CW laser diodes described in the embodiment 2, in the external sample of all thermal release horny layer that makes according to embodiment 1, all observe sure ablation result, but wherein some are better than other.Especially CuPc (CPC) and epolite #67 list in the most effective among.The possible explanation that CPC has outstanding performance is that its high boiling point that has more than 500 ℃ still was in solid phase with it before reaching this temperature.

Embodiment 5

Because CuPc is allowed to use in transplantable suture by U.S. food Drug Administration and is listed in " Merck index " as quite optimum and stable molecule aspect the biological compatibility of people, the next step of being taked is that the local application on the volunteer skin of health combines with CPC and focused light source.The suspension of preparation CPC fine powder in isopropyl alcohol.The application process that adopts is to sway solution, applies a droplet at target site then.Along with the vaporization of alcohol, stay the thin of solid phase CPC and uniform coating at skin surface.

Then device shown in Figure 1 is applied on the skin local the coating on the above-mentioned position of CPC, with the selection area of subject's skin facing to datum plate.Datum plate is had the thin windowpane hole, about 3cm * 3cm of 4mm size to form by the center.Then the zone that CPC is covered is placed within the centre bore.Then confocal video microscope (Fig. 1) is clearly focused on skin surface.Placed can obtained in video system on the position of the most clear focus, even the focus of laser system overlaps with skin surface.The operator starts laser pulse then, simultaneously with the effect on the videomicroscopy observation target site.Increase along with the micropore degree of depth, the operator is by the defocus amount of Laser Measurement point in micropore, the amount of penetrating is carried out visual assessment, and along with skin histology is goed deep on ablated surface, the operator can move forward into the action attitude along " z " axle by the position with camera/lasing light emitter and proofread and correct.When horny layer was removed to epidermis, the appearance significant change of bottom, hole became more moistening, more glossy.After observing this variation, the operator can disconnect laser.In many cases, according to hydration status and other physiological conditions of trier, as the result that the cuticular barrier function on this zonule is removed, interstitial fluid significantly outflows.Write down this visual record of the interstitial fluid accessibility of site of perforation with video system.

Embodiment 6

Repeat the method for embodiment 5, different is that CPC is applied on the transparent adhesive tape, this adhesive tape is sticked on the selected position of subject's skin then.Basic identical among its result and the embodiment 5.

Embodiment 7

By method well known in the art corpse skin is carried out histology experiment to determine the ablation threshold parameter of the dye mixture that provides and relevant damage information.End face with the solution-treated skin sample of CuPc (CPC) in alcohol.After treating alcohol vaporization, the partial layer of solid phase CPC just is distributed on the skin surface, its average thickness is the 10-20 micron.Fig. 8 A is the cross section of the skin of full depth before laser is used, and wherein 270 is CPC layers, the 274th, and horny layer, the 278th, following epidermal area.Fig. 8 B uses the single light pulse of 810nm with 4000W/cm ²Energy density and pulse period of 20 milliseconds be applied in 80 microns of diameters circle in after sample.Even it should be noted that central authorities, still have the CPC of significant quantity to be present in cuticular surface at ablation swallow 282.The result who should also be noted that laboratory measurement shows that being incident on only has 10% by actual absorption in the luminous energy on the CPC, and other 90% are reflected or backscatter.Therefore, thus transfer to the only about 4000W/cm of effective energy flux that dye coating can cause required heating ²Fig. 8 C is 5 light pulses sample afterwards of using 810nm, and wherein the cutin barrier layer is removed and following tissue is not had any damage.These have represented the thermal ablation performance that " ideal " optics is regulated dry straightly.Fig. 8 D uses 50 subpulses sample afterwards.Because the thermal denaturation of the carbonization of ablation tissue and following tissue damaged tissues 286 do not occur at epidermal area.Fig. 8 A-8C has shown separating between horny layer and the following epidermal area, and this is owing to dehydration, freezing and prepare the breast that imaging occurs and resemble (artifact).

Embodiment 8

For getting the details of thermal ablation mechanism clear, set up the mathematical model of skin histology, can attempt the various embodiment of thermal ablation method at this model.Temperature Distribution in this Model Calculation multilamellar semo-infinite medium is imported given heat flux on surface local ground, remove from the surface of the segment distance of being separated by and reduce phlegm and internal heat, and promptly between carries out convection current.In cylindrical coordinates, solve axisymmetric time dependent diffusion equation with implicit expression alternating direction (ADI) method.(annotate: use the constant temperature boundary condition as z → infinity at lower boundary; Making radially on the maximum radial border, heat flux is zero as r → infinity).Each layer is parallel with skin surface, and they are respectively (1) dyestuff; (2) horny layer; (3) Xia Mian epidermis; (4) corium.For each layer, the degree of depth in the semo-infinite medium and hot property, density (rho), specific heat (c) and conductivity (k) all must be determined.

At first, calculate the heat transfer coefficient h of skin according to " stable " determined by ambient air temperature, skin surface temperature and dermal temperature, " one dimension (1-D) " Temperature Distribution.Supposing at skin surface does not have the dyestuff existence and provides " h ".Program can allow the technical staff use on the dye coating surface and be somebody's turn to do " h " or import another required " h " for the dyestuff surface.Then, " h " that determines with the dyestuff surface calculates " stable " Temperature Distribution to all layers (comprising dye coating).This Temperature Distribution is the initial condition with the heating problems of time correlation.This has constituted " m-file " initial m.The temperature field of program by passing, calculating and each step of demonstration of time solves the Temperature Distribution with time correlation then.

Each embodiment of method described herein (their empirical data is collected) has carried out modelling with regard at least one group of operating parameter, and its shows that horny layer is ablated can be accurately and the controllable mode realization.Simulation output illustrates with two kinds of different forms: (1) has shown the viewgraph of cross-section of the skin of different tissues layer, at this 3 isothermal lines is arranged above the view, and they have provided 3 critical temperature threshold values; Article (2) 2, the curve of different temperature and times relation, a point that is illustrated in the horny layer central authorities under the target site, another is illustrated in the activated cellular layer of epidermis and the borderline point between the cuticular bottom surface.How the temperature of these curve display each point when applying thermal pulse as technical staff in tissue has implanted micro thermocouple along with the time changes.In addition, use this model can study the limit of parameter, this method can be in order to the outer limits of two importances determining this method performance in this limit range.The explanation situation that can define boundaries at first can be used this method and not cause pain or undesirable tissue injury in the boundary line.

As described at several different embodiments of the present invention, for any given thermal source, exist a effect to become and be not best point subject's skin histology, because the subject feels pain, perhaps because activated cell is subjected to temperature action in following epidermis and/or the corium, and this is if keep the sufficiently long persistent period, will cause the damage to these tissues.Therefore, local CuPc (CPC) dyestuff embodiment with the optics heating carries out test simulation as base-line method, with the hot thermal constant of establishing different skin histology layers is how to determine a window basically, can use the method and do not have pain or adjacent organized layer is produced damage in this window.

Fig. 9 and 10 is cross sectional representation of skin layer and local dye coating.In each figure, shown 3 isothermal lines clearly: (1) 123 ℃, it is the temperature that water vapor in the tissue causes tissue ablation; (2) 70 ℃, if this temperature continues the several seconds, activated cell will sustain damage; (3) 45 ℃, the subject will feel the mean temperature of pain.This pain threshold has narration in several basic physiology's textbooks, but experience shows that this threshold value is subjective factors more or less.In fact, in the repeated trials that same subject is carried out, only be to form different holes in several millimeters the scope, can produce significantly different sensation of degree, this may be because due to the contiguous teleneuron of site of perforation.

Dimension on the curve chart shows the dyestuff measured with micron and the different layers of skin, between them with smooth border separately.Yet the border of actual skin histology far is complicated, and on average meaning, for related dimension, this model has provided good being similar to of the thermal gradient that exists in the actual tissue.In all simulations of this simulation and back, the dimension of CPC dye coating and each skin layer is as follows: dyestuff, 10 microns; Horny layer, 30 microns; Following epidermis, 70 microns; Corium, 100 microns.

To the additional conditions of the model that is used for this physical simulation table of face as follows:

Table 1

The initial condition of difference thermal model
		Ambient air temperature	Ta＝20℃
Skin surface temperature	Ts＝30℃
		Dermal temperature	Td＝37℃
The dyes vaporize temperature	Tvap＝550℃
		The horny layer vapourizing temperature	Tc1＝123℃
Tissue injury's temperature	Tc2＝70℃
		" pain " temperature	Tc3＝45℃
The radius of irradiation area	R hot=30 microns
		The energy density of using	FLUX＝400W/cm 2

Table 2

Parameter	Dyestuff	Horny layer	Epidermis	Corium
					Thermal conductivity	0.00046	0.00123	0.00421	0.00421
Density	0.67	1.28	1.09	1.09
					Specific heat	0.8	1.88	3.35	3.35

When these simulations of utilization, additional following careful supposition:

1. when a part of horny layer may have the temperature of the ablation threshold that surpasses the vaporization of moisture heat, this incident was not included in the model, and the heat-energy losses that occurs in tissue subsequently owing to this vaporization does not turn to a factor in the above-mentioned simulation yet.Temperature shown in the tissue during this will cause simulating below from this moment slightly raises.

2. similarly, when a part of CuPc (CPC) dye coating had reached its evaporation point of 550 ℃, this incident was not included in the model yet, but only temperature is limited in this level rigidly.Carry out along with mimic, this also will cause the following temperature subsequently of organizing slightly to raise.

Although used these reduced conditions in the model, the performance of expection and according to clinical research with to the Histological research of donor tissue sample and the relatedness between the observed by rule of thumb performance is significant.The critical data that notes in 10 at Fig. 9 is the position that applies 3 shown different threshold temperatures of the time span of thermal pulse and isothermal line.

In Fig. 9, pulse length is 21 milliseconds, and 70 ℃ of isothermal lines are just in time crossed the border between the activated cell in horny layer and the epidermis.Under this condition, the in vitro tests that donor skin sample is carried out shows that 50 apart 50 milliseconds heat pulses can detectedly damage (seeing Fig. 8 D) to this top layer generation of living cells.Yet in vitro tests shows that also under these same operation parameters, 5 heat pulses are organized these and do not produced significant damage.So can think that even surpassed nominal damage threshold (at least on the meaning of a transient state), this temperature must totally keep certain incident, just can produce damage by real pair cell.But the essential information that obtains by simulation is that if remain on below 20 milliseconds " turn-on time " of thermal pulse, and flux density is 400W/cm ²Even ablation threshold isothermal line has goed deep into horny layer, the living cells in the epidermis in its lower section can not sustain damage yet.In other words,, regulate suitably shortly, just can realize cuticular ablation and can not damage contiguous cell (seeing Fig. 8 C) in the epidermis below it its " turn-on time " by using the thermal source of small throughput density.This may mainly be because these two kinds of layer tissue layers have significantly different heat diffusivity.That is, the thermal conduction constant 0.00123J/ (ScmK) of horny layer (only containing the 10-20% moisture of having an appointment) is more much lower than the thermal conduction constant 0.00421J/ (ScmK) of epidermis.This just can make the temperature in the horny layer improve gradually, and (intensification) remain in the strict spatial dimension, until ablating.

In Figure 10, same simulation is begun further to carry out from damage threshold critical point computing shown in Figure 9.By in the circle of 60 microns of the diameters of heated dyestuff with same flux density 400W/cm ²Apply 58 milliseconds of thermal pulses, just in time enter the weak layer of the skin that is comprised by corium at 45 ℃ pain perception isothermal lines.In addition, the damage threshold isothermal line is more deep significantly to epidermal area in the position shown in Fig. 9 than it.This simulation is connected with numerous clinical researches of carrying out with the method, and the accuracy of model obtains fabulous confirmation because models show almost be exactly before the subject feels thermal probe can be applied in and continue " turn-on time " on the skin.In clinical trial, set " turn-on time " and " turn-off time " of a series of light pulses on CuPc (CPC) partial layer that is applied on the skin with the controllable pulse generator.When keeping constant " turn-off time " of 80 milliseconds, " turn-on time " progressively increases up to the subject and tells slight " pain " sensation.Without exception be that all subjects in these researchs tell " pain " for the first time in " turn-on time " for time between the 45-60 millisecond, and are desired very approaching with model.In addition, in these clinical researches, noticed the situation that foregoing " pain " sensation becomes with the position.Therefore, tell to be the point that can discover for the first time clear and definite sensation for " pain ".At a position, may tell to be pain, and in adjacent regions, same subject may tell to only " discovering ".

A key element of this clinical research is comprehension, even at same position, the impulse train of the non-homogeneous of thermal pulse can work with subject's psychological physiological neural consciousness one, causes that the actual of sensation go down.For example, the short pulse of available a series of length makes the neurocyte in the zone saturated, temporarily makes the neurotransmitter depletion in this synapse, thereby limits the ability of its transmission " pain " information.Consequently, this order long its beginning in this sequence of pulse ratio after these short pulses is used and is difficult for discovering.Therefore, carried out a series of experiments with some impulse trains of making arbitrarily, the gained result is consistent with this hypothesis.This saturated similar situation can be found in the sensation when following situation: when a people had just begun to step into very hot bathing pool, beginning had pain perception, but along with he adapts to the sensation of heat, just promptly became and can stand.

Embodiment 9

An object of the present invention is to realize cuticular painless micropunch and can not produce significantly to damage to activated tissue on every side.Described in the simulation of explanation in embodiment 8 and Fig. 9-10, as if the heat energy for a given flux density in the ablation target spot exist strict border, and painless AT mode that just in time can be such in this border is carried out micropunch.All shown such situation with in vitro study in the body, this makes that method is obtained the certain operations parameter by experiment, and these parameters can obtain good result.Following mimic combination shows the effect of the method when using these concrete parameters.

In first kind of situation, will be applied in by an impulse train of 10 pulses on the skin of CPC covering, " turn-on time " respectively is 10 milliseconds with " turn-off time ".Figure 11 has shown that the final temperature in the skin histology after this impulse train has just finished distributes.Can see that the isothermal line of three critical temperature threshold values of expression shows has realized that horny layer ablates and the neural no any sensation of skin corium, and damage threshold enters in the following epidermis activated cell seldom.As previously described,, not only epidermis cell must be heated to a certain temperature, but also they must be kept a period of time in this temperature, it is generally acknowledged about 5 seconds for the persistent damage of the actual generation of pair cell.Figure 12 and 13 has shown that respectively the temperature (as time function) of horny layer and activated epidermis is in " turn-on time " heating with in " turn-off time " refrigerative whole 10 circulations.Interrelate studying in this simulation and the body that carries out, notice in the usefulness more than 90% and set in the perforation trial of carrying out with the corresponding to systematic parameter of above-mentioned simulation, realized cuticular effective perforation and the subject does not have painful sense, in the microscopy of the site of perforation of after a few days, carrying out, be not checked through obvious damage to tissue.The result of the in vitro study that the donor skin sample of whole thickness is carried out is consistent with the model expection also.

Embodiment 10

In the body that carries out, in experimentation and these simulations, seem that the pre-cooled of skin helps to reduce pain or surrounding tissue is produced the probability of damage, makes the micropunch process optimization.In practice, this can easily be realized by following method: before perforation, be close to simple coldplate of placed.For example, in the circle of 1 centimetre of the diameter around the perforation target site, use Peltier (Peltier) coldplate (plate keeps several seconds at about 5 ℃), can significantly reduce the temperature of tissue.Fig. 3 A-B is the sketch map that is used for this purpose experimental provision in the laboratory.By will with identical 10 cycle pulses that use in the computing of explanation among the embodiment 9, can find desirable much improvement aspect the temperature length of penetration of going deep into skin histology is controlled after Figure 11 and Fig. 4, Figure 12 and Figure 15, Figure 13 and Figure 16 compared.Have, compare with corium with epidermis, cuticular lower thermal diffusion coefficient and specific heat are favourable.In case cooling, the tissue of the high degree of hydration of epidermis and corium need bigger heat energy input promoting its temperature, and cuticularly organize more driedly, can be heated to ablation threshold rapidly.

Embodiment 11

In case understood with power transfer to skin histology to carry out the basic thermal conduction mechanism of painless ablation of effective horny layer and micropunch, can visualize several different concrete grammar of realizing that required contact point fast temperature is regulated, for example the hot wire embodiment of explanation in Fig. 4-7.

As described here, a kind of basic embodiment is to use Ohmic heating element (Fig. 4), the tip of for example little cordless soldering iron is wound with sizeable, inactive relatively tinsel around the Ohmic heating element, remaining short tinsel is outstanding from heater device body.When applying electric current with constant current source, heater will be increased to a certain temperature, and in seconds by convection current towards periphery the air release heat reach stable state.Similarly, also will reach stable state as the tinsel of this hot system part, like this, when using such parts, tip wiry can rise to almost any temperature arbitrarily, up to about 1000 ℃.The size of this tip can be made that can to provide just in time be required micropore size.

In laboratory, used the tungsten filament of 80 microns of diameters, be connected on the replaceable tip of " WAHL " cordless soldering iron, tinsel protrudes about 2mm from tip.With the temperature of thermocouple measurement tip in stable state, notice by changing the value of setting of constant current, can easily reach the steady temperature more than 700 ℃.For realizing required adjusting, the electro-mechanical actuator and the tip of low quality fast-response linked, like this, position wiry can be up to the speed of 200Hz linearly more than the translation 2mm.Then, by whole device is fixed on the precision surface plate, the tip of this vibration is very controllably contacted with skin surface: tip is only in each " turn-on time " and contact skin below 10 milliseconds, by correspondingly setting " turn-off time " that pulse generator then can reach random length.Studies show that in these bodies, can before perforated subject knows tinsel tip and contact skin, realize perforation really.

For the performance of present embodiment and the Local C PC dyestuff embodiment of optics heating are compared, carry out following simulation by the method for embodiment 8.Basically only by changing initial condition, available same simulation code carries out the hot wire embodiment.Because with wiry the contact is instantaneous basically, heat can be along with the variation accumulation of time in the CPC layer, when tinsel is physically removed with the contacting of skin, there is not any residual heat from the teeth outwards, and residual heat is arranged on the CPC dye coating that heated, also have,, before it is applied in horny layer, should not have the horizontal proliferation of any heat energy because tinsel self has been determined the target region of ablation/micropunch.The comparison performance of " hot wire " embodiment is seen Figure 17-19.

Embodiment 12

In the present embodiment, use the method for embodiment 11, different is to carry out pre-cooled by the method for embodiment 10 to skin.Similarly, in " hot wire " embodiment, to the similar useful result of the pre-cooled generation of target site.The pre-cooled Simulation result of " hot wire " method is seen Figure 20-22.

Embodiment 13

As if as what discussed in the background introduction of this paper, TankovichShi ' 803 patents are similar to claimed invention herein at first view.Originally executing in the example, using the operating parameter of defined in TankovichShi ' 803 patents to make up analogue model, promptly pulse width is 1 microsecond, and power level is 40,000,000W/cm ²Figure 23 and 24 shows, the horny layer that does not have any part under these conditions reaches promptly 123 ℃ of the threshold values of water flash distillation, therefore, horny layer ablation/micropunch does not take place.In practice, the high-peak power of this type, the pulse of short persistent period are applied on the local dye coating, only make the dyes vaporize of skin surface and skin is not had any effect.Therefore, present embodiment proves, the condition of TankovichShi ' 803 patent defineds is not render a service in the claimed invention herein.

Embodiment 14

In the present embodiment, the interstitial fluid that collection obtains after boring a hole on skin by the method for embodiment 6, and analyze to measure its concentration of glucose.Data are not suffered from the subject of diabetes and the subject of 6 trouble diabetes of just carrying out glucosieloading test obtains from 4.Subject's age is at 27-43 between year.The target of this research is the purposes of research the method aspect following: collect enough interstitial fluids (ISF) on one's body from the subject painlessly and can carry out the glucose content analysis to make the ISF sample, then the concentration of glucose in these concentration and the subject's whole blood is compared.

" ELITE " system that all subjects' blood and ISF all used Miles-Bayer company has carried out glucose analysis.All 10 subjects adopt same assay method, but those patients that suffer from insulin-dependent diabetes are being adjusted aspect glucose load and the insulin input.

The basic design of this research is the volunteer of enlisting right quantity (some of them suffer from diabetes and other do not suffer from diabetes), in during 3-4 hour the whole research every 3-5 minute right from the sample of a series of ISF of these volunteer withdraw and whole blood.Blood and ISF sample are all carried out glucose analysis, determine the statistics relation of blood sugar level and interstitial fluid.For studying the temporary sluggishness of comparing the hypothesis of ISF glucose level with the full blood glucose level, induce the subject on its glucose level, to show significant dynamic change.This realizes by following method: before beginning test, allow each subject's fasting 12 hours, make its baseline glucose level allow it carry out glucose load after establishing by 3 in cover blood and ISF glucose level on an empty stomach then.After baseline values is established, allow the subject carry out glucose load with the form of hesperidium juice according to following policy:

I. for the subject of matched group, according to the criterion calculation glucose load of 0.75 gram glucose/weigh oneself.

Ii. for the subject who suffers from insulin-dependent diabetes, glucose load is 50 gram glucoses.In addition, after carrying out glucose load, the subject who suffers from diabetes with the insulin of the snap action of the dosage in its of self-injection immediately in normal morning.Under the subject's who suffers from diabetes the situation of glucose level on an empty stomach at 300mg/dl, they are required first self-injection insulin, fall after rise in its blood sugar level and carry out glucose load to 120mg/dl.

In " sensible agreement " file, give the explanation fully of a research of each subject of enlisting earlier, require them before formal participation program, to understand and signature.After the acceptance, they promptly fill in history questionnaire.The concrete clinical method of implementing is as follows:

(a) subject can only drink water since 9 extremely research fasting in evening.Do not allow to drink coffee during this period, fruit juice or smoking.

(b) subject arrived test mechanism before 9 of the next mornings.

(c) subject falls to being seated in the oblique chair, and this oblique chair provides to the subject so that it loosens in whole research process.

(d) after the subject arrives, promptly begin to extract whole blood and ISF sample, and in ensuing 3-4 hour, proceed every 3-5 minute.Collect data the duration according to glucose load after when subject's blood sugar level return to and also decide normal range.With below the optics perforation method of describing in detail (the ISF pump is inhaled method) being collected the ISF sample.The volume of each ISF sample is about 5 microlitres to guarantee the filling ELITE test tape well.Take each blood sample with finger lancet commonly used.ELITE domestic glucose meter system with Miles-Bayer company carries out glucose analysis to ISF and blood sample immediately.For improving estimation, each thorn is referred to that sample carries out two independently ELITE analyses to " very " blood sugar level.

(e) become easy for the ease of collecting ISF continuously from given subject's same position in whole data collection phase, on forearm on the subject, punctured and constituted 25 micropores of 5 * 5 matrixes, the width of each micropore is between the 50-80 micron, each other at a distance of 300 microns.Diameter 30mm, center are had the polytetrafluoroethylene disk in the hole of a 6mm size be bonded at subject's forearm with contact adhesive, the position of centre bore that makes the 6mm size is in the top of 5 * 5 rectangular micropores.Because this is bonding, thereby can adopt a kind of like this short-cut method: connect little suction pipe, use medium vacuum (the 10-12 inch of mercury) in punched areas, ISF is flowed out in body by micropore.Transparent windowpane is equipped with at the top of polytetrafluoroethylene disk, but makes below operator's direct observation the skin of micropunch.When the 5 microlitre drops of ISF form on skin surface, can easily with the naked eye determine by this window monitoring micropunch position.The vacuum of this degree produces 5 pounds/inch ²(PSI) nominal pressure gradient.There is not micropore, in any case be can't extract ISF on one's body out only from the subject with medium vacuum yet.

(f) extract out three samples to after, drink high sugared Fructus Citri tangerinae juice to the subject, allow it carry out glucose load.For the subject who does not suffer from diabetes, the glucose amount that gives is 0.75 to restrain/weigh oneself, and the subject who suffers from diabetes then is 50 grams.The subject who suffers from diabetes goes back the insulin of (periodically) self-injection potion (daily) snap action, and its dosage is taken into account the picked-up of glucose load according to this 50 gram glucose level and suitably calculated.For throwing in insulin to normal 1.5-2.5 hour sluggishness between the maximum efficiency of medicament, the ascensional range that subject's blood sugar level of diabetes is suffered from expection can reach 300mg/dl, falls after rise to normal range after insulin works then.Expect that the subject who does not suffer from diabetes presents the glucose tolerance test result of markization, usually occur the peak (between 150-220mg/dl) of blood sugar level in 45-90 minute after carrying out glucose load, fell after rise to its normal baseline values rapidly at ensuing about one hour then.

(g) carrying out after glucose load or glucose load add injection of insulin, in ensuing 3-4 hour, extracting subject ISF sample simultaneously and thorn refers to whole blood sample with 5 minutes interval. when showing that subject's glucose stablize, the blood sugar level in 3 continuous samples stops taking a sample.

After checking data, can obviously see several characteristics.Especially, for the ELITE test tape of any particular batch, compare with the level that shows in the blood, there is tangible skew in the output that shows with the mg/dl glucose in the glucose meter.Owing in ISF, do not have erythrocyte (hematocrit) also owing to the electrolyte difference between ISF and the whole blood, the reading that expection can obtain increasing.No matter what the fundamental cause of output offset is, by with reference to the comparison of analyzing, determine real ISF glucose level and the numerical value that obtains by the ELITE system between linear, it is constant that the ELITE of any particular batch is with its proportionality coefficient.As a result, for ISF glucose level and whole blood measurement result are compared, the ISF data are carried out following one-level linearity correction: ISF _Glucose=0.606*ISF _ELITE+ 19.5.

When being used for measuring the ISF G/W at ordinary times, the conversion of this ELITE output makes the technical staff can be to the whole data set inspection error term relevant with using ISF to assess blood sugar level.Certainly, even do not do any linear conversion, dependency between ISF dextrose equivalent and the blood sugar level and conversion type identical.

According to literature content of delivering about the majority of ISF glucose aspect and preliminary data, be expected at originally between the glucose level in the whole blood that ISF glucose level and cluster needling refer to obtain and observe 15-20 minute sluggishness.Result displayed when this is not analytical data.Specifically, when the data set of analyzing each subject when determining to obtain the required time migration of maximum correlation between ISF glucose level and the blood sugar level, find under the situation the poorest among this group subject, time lag only is 13 minutes, average time, sluggishness only was 6.2 minutes, and several subjects' appearance almost are (about 1 minute) time responses immediately.

According in the observed minimum sluggishness of this data set, curve chart shown in Figure 25 has provided all 10 glucosieloading tests, they expansion the time put on one and in succession one.The data that provide do not have skew any time, show that the high level between ISF and the blood sugar level is followed the tracks of, and whole clinical data group is handled with identical mode.If whole data set is offset as a whole to find best estimator time response, the dependency between ISF and the blood sugar level reaches peak value when r=0.97, and be 2 minutes time delay.This only is the insignificant improvement that the unmigrated dependency by r=0.964 carries out.Therefore, for the analysis of remainder, their ISF value is handled on not additional skew any time ground.That is, blood and ISF glucose level are respectively organized in the same processing with the data of collecting simultaneously.

In unmigrated Elite ISF reading being converted, after the ratio of glucose, can check the error relevant with these data with reflection ISF.The simplest method is that two ELITE thorns of supposition refer to that glucose readings in fact is the value that is absolutely correct, and ISF value and these average blood sugar values after only will converting then compare.These data are specific as follows: the standard deviation of blood-ISF, 13.4mg/dL; The coefficient of variation of ISF, 9.7%; The standard deviation of two Elite, 8.3mg/dL; The coefficient of variation of blood (Miles company provides), 6%.

Shown in these data, be that basic measured value has contained error term with blood.In fact, the performance data that manufacturer delivers shows that the nominal coefficient of variation (CV) of ELITE system depends on erythrocytic amount in the gentle blood of G/W between 5-7%.

Another style of difference is presented among Figure 26 with the form of scatter diagram between ISF glucose and the blood glucose.In the figure, the upper bound of 90% confidence interval and lower bound have also been shown for your guidance.It should be noted that all data in the scope of blood sugar level below 100mg/dL all fall within these 90% confidence interval error bars, only have two exceptions.This is very important, because do not measure the tendency of glycopenia, will bring very serious consequence to the user who suffers from diabetes.That is, cross the lowland glucose level of forecast between 40-120mg/dL and compared the highland forecast better.

Basically, if the analytical error of the fundamental analysis error when supposition is used the ELITE system to ISF when whole blood is used the ELITE system can be compared, then the ISF glucose can be expressed as follows the deviation of blood glucose:

ISF _Deviation=((ISF _Actual) ²+ (blood _Actual) ²) ^1/2

The numerical value that this equation is applied to provide above, can find the solution " truly " value of the estimation of ISF error term:

ISF _Actual=((ISF _Deviation) ²-(blood _Actual) ²) ^1/2

Perhaps, in order to following equation solution:

ISF _Actual=((13.4) ²-(8.3) ²) ^1/2=10.5mg/dL.

ISF is shown in Figure 27 to the rectangular histogram of the relative deviation of blood sugar level.

By the hole administration in horny layer

The present invention also comprises a kind of method of carrying medicine (medicine that comprises present percutaneous conveying) by the micropore in the horny layer.In an illustrative embodiment, realize administration by the top reservoir that solution is placed site of perforation.In another illustrated embodiment, further promote administration with barometric gradient.In another embodiment, further promote administration with ultrasonic energy, can use or working pressure gradient not this moment.The operation of ultrasonic energy can or utilize acoustic streaming effect (will be explained) so that the horny layer of the solution of carrying by boring a hole according to traditional percutaneous parameter at once.

Embodiment 15

Present embodiment shows by bore a hole delivery of lidocaine (a kind of local anesthetic) at horny layer.Lignocaine solution also contains chemosmosis promoter preparation, and it is that design promotes lignocaine to see through cuticular passive diffusion.The figure of illustrative conveyer device 300 sees Figure 28, and wherein this device comprises the shell 304 around reservoir 308, and reservoir is used for containing and contains drug solns 312.The top of shell comprises ultrasonic transducer 316, and it can provide ultrasonic energy to help will to contain drug solns by micropore 320 administrations in the horny layer 324.Can the openning 328 from ultrasonic transducer exert pressure and contain drug solns by the micropore administration in the horny layer with further help.Conveyer device is applied in a selection area of subject's skin, device is placed at least one, on best a plurality of micropores.The adhesive phase 332 that is attached to outer casing underpart can make device stick on the skin, like this, and containing drug solns and can flow in the reservoir from micropore.Result by the micropore administration is that medicine inputs in following epidermis 336 and the corium 340.

With 5 subjects the effectiveness of utilizing perforation and ultrasound wave administration is tested.Carry out at two positions of experiment on subject's left forearm, and two positions are separated by about 3 inches, are symmetrically located between thumb and the upper arm.Position near thumb is called position 1, is called position 2 from thumb position farthest.Position 1 is used lignocaine and accelerator solution with same conveyer device 300 at this position as the contrast position, but not at horny layer middle punch or use ultrasonic energy.24 holes of puncture in the circle of 1 centimetre of the diameter at position 2, these holes at a distance of 0.8 millimeter, are grid type each other.The method of embodiment 6 is adopted in 2 perforation at the position.Use lignocaine and low-level ultrasound wave.Hyperacoustic using is to use Zevex type ultrasound transducer means customized, be set to pulsed, peak-peak to ENI # 2100L linear amplifier is input as 0.4 volt, can produce 1000 short pulses that occur with 10Hz, fundamental frequency is 65.4KHz,, obtains pulse modulated signal that is, transducer by energy excitation, is then turned off 85 milliseconds in 15 milliseconds of short pulses.The amplifier that records is output as 0.090 watt of RMS to transducer.

Use after the lignocaine, rub back and forth at the test position, measure sensation with No. 30 tinsels.Experiment is carried out at two positions, and 1 carried out 10-20 minute at the position; At the position 2 successively carry out two 5 minutes.The numb degree at two positions of level evaluation with 10 to 0,10 expression subjects do not have feeling of numbness, and 0 expression subject feels complete numbness.Be all 5 subjects' synthesis result below.

The contrast position is that position 1 was 10-12 minute very little and even nothing (grade 7-10) of feeling of numbness.At about 20 minutes, along with solution permeates horny layer fully, 1 observed some numbness (grade 3) at the position.Use the back that finishes at lignocaine and clean position 1.Position 2 is almost completely numb (grade 0 to 1) in the circle of 1 centimetre of the diameter with micropore.Outside the circle of 1 centimetre of diameter, feeling of numbness almost straight line descends, and is 1 at 2.5 centimetres circles, does not then have feeling of numbness outside 2.5 centimetres circle.The result of the assessment at the position 2 after using for the second time is, complete numb diameter of a circle is bigger, is about 1.2 centimetres, and feeling of numbness drops to 1 with irregular oval pattern straight line, oval-shaped 2-2.5 centimetre diameter is perpendicular to forearm, and its 2-6 centimetre diameter is parallel with forearm.Outside this zone, do not observe numbness.The pictorial representation of the illustrative result that obtains on one's body representational subject is seen Figure 29 A-C.Figure 29 A and 29B have shown the result that 2 (perforation) obtain at the position after 5 and 10 minutes respectively.Figure 29 C has shown the result that 1 (contrast position, puncherless) obtains at the position.

Be used to promote mobile ultrasonic energy of percutaneous and promoter

Can utilize physical action in one approach, by this method, can modulate the flow velocity that reaches with additive method to improve audio frequency by the ultrasonic energy field of ultrasonic transducer generating.Shown in Figure 1 as U.S. Patent No. incorporated by reference herein 5,445,611 can be divided into the Energy distribution of ultrasonic transducer near field and far field.Near field (characterizing with length N) is the zone from first energy minimum to last energy maximum.The zone that is distal to last maximum is the far field.Closely (N) field mode comprises local pressure peak and the null value that a large amount of intervals is very near.The length N near field is the function of speed that the shape of frequency, energy converter planar and size and ultrasound wave are advanced in medium.For single transducer, the Strength Changes in its normal operation range can't influence the character that ultrasonic energy distributes, except with linear mode.Yet for the system with a plurality of transducers (all frequency and amplification being modulated), the relative intensity of independent transducer can influence the Energy distribution in the acoustic medium, no matter medium is skin or another kind of medium.

By the frequency of appropriateness (for example between about 1-20%) change ultrasonic energy, it is relative constant that the pattern of peak and null value keeps, but the length N of near-field region changes with frequency with being directly proportional.The great change of frequency (for example more than 2 times or 2 times) will be probably produces not resonance or vibration mode on the same group in transducer, cause unpredictable significantly different near-field energy pattern.Therefore, for the moderate change of audio frequency, the complex patterns of peak and null value will compress or expand in accordion shape mode.By selecting warbled direction, the offset direction at these local pressure peaks of may command.By at skin surface ultrasound application energy, these local pressure peaks of the selectivity of audio frequency modulation may command move towards the inside of health or towards the surface of health by skin.Modulating frequency can be ordered about pressure spike and entered in the body from high to low, and modulating frequency is then pushed pressure spike body surface to and pushed to external by skin in body from low to high.

Adopt the canonical parameter of this method of application, for example, the ultrasonic transducer diameter is 1.27 centimetres, and the nominal operation frequency is 10MHz, acoustic impedance is similar to water, and then the frequency modulation(PFM) of 1MHz makes the peak of near-field energy pattern and null value produce moving of about 2.5mm near horny layer.From the angle that the percutaneous of analyte and/or through mucous membrane extract, the acting as of this degree enters the zone below the horny layer, or even its hetero-organization below epidermis, corium or the corium provides good methods.For any given transducer, may there be the optimum frequency scope, frequency modulation(PFM) is the most effective in this scope.

Also can improve medicine or the percutaneous speed of analyte by changing resistance (diffusion coefficient) or driving force (diffusion gradient).Using so-called penetration enhancer is that chemical promoter can improve speed.

Chemical promoter is grouped into by the one-tenth of two kinds of main types, is promptly upset chemical compound and solvent or is contained the binary system that cell membrane is upset chemical compound and solvent by cell membrane.It is known in this area that cell membrane is upset chemical compound, and known its can be used for the preparation of local application's agent, also works in by skin extraction analyte.These chemical compounds are considered to help dermal osmosis by the lipid conformation disordering that makes the corneocyte film.The european patent application No.43 that the catalogue widely of these chemical compounds is incorporated by reference herein, 738 (date of publication: June 13 nineteen eighty-two) narration is arranged.It is believed that any cell membrane upset chemical compound all can be used for the present invention.

Suitable solvent comprises water, glycol (for example propylene glycol) and glycerol; Monohydric alcohol (for example ethanol, propanol) and higher alcohol; DMSO; Dimethyl formamide; N,N-dimethylacetamide; 2-Pyrrolidone; N-(2-ethoxy) ketopyrrolidine; N-Methyl pyrrolidone; Alkyl azacycloalkyl-2-ketone (azone) that 1-dodecyl-aza-cycloheptane alkane-2-ketone and other N-replace etc.

The U.S. Patent No. of announcing on August 27th, 1,985 4,537,776 has comprised the outstanding summary of prior art and background information, describes the use that is used to promote some binary system of permeating in detail.Because this announcement is comprehensive, information and term that this article uses are incorporated by reference herein.

Similarly, above mentioned european patent application No.43,738 have pointed out how to use the selected glycol and the cell membrane of numerous species as solvent to upset chemical compound and carry the lipotropy pharmaceutically active compounds.Because it has at length disclosed cell membrane and has upset chemical compound and glycol, the content that discloses in this european patent application is also incorporated by reference herein.

The UK Patent Application GB 2 that on August 21st, 1985 announced, 153,223A has disclosed a kind of binary system that is used to promote the metoclopramide infiltration, and it is by C8-32 aliphatic monocarboxylic acid's (undersaturated and/or branched (if carbon number is 18-32)) monovalence alcohol ester or C6-24 aliphatic monohydric alcohol (undersaturated and/or branched (if carbon number is 14-246)) and N-cyclic compound (for example 2-Pyrrolidone, N-Methyl pyrrolidone etc.) formation.

U.S. Patent No. 4,973,468 disclose, and the promoter combination of being made up of carbitol or monomethyl ether and PGML and methyl laurate can promote steroid such as percutaneous dosings such as progestogen and estrogen.The dual promoter of being made up of glycerol monolaurate and ethanol that is used for the medicine percutaneous dosing is disclosed in U.S. Patent No. 4,820, in 720.U.S. Patent No. 5,006,342 have listed many promoter that are used for percutaneous dosing, and they are by fatty acid ester or C ₂To C ₄The aliphatic alcohol ether of alkanediol is formed, and wherein each fatty acid/alcohol moiety of ester/ether contains 8-22 carbon atom.U.S. Patent No. 4,863,970 disclose the compositions of the promotion infiltration that can be used for local application, these compositionss comprise the carrier that promotes infiltration, and one or more cell membrane that described carrier then contains active penetrant and specified amount are upset chemical compounds (for example oleic acid, oleyl alcohol and oleic glycerine ester; C ₂Or C ₃Alkanol and inert diluent such as water etc.).

Other chemical promoters (might not be relevant with binary system) comprise that Herschler is in U.S. Patent No. 3,551,554,3,711, DMSO that points out in 602 and 3,711,606 and the aqueous solution of DMSO and azone (alkyl azacycloalkyl-2-ketone that N-replaces and for example Cooper in U.S. Patent No. 4, the chemical compound of pointing out in 557,943).

Some chemical promoter may have bad side effect, as toxicity and skin irritation etc.United States Patent (USP) 4,855,298 have disclosed a kind of like this compositions, and it contains the glycerol that presents in an amount at least sufficient to produce the irritation effect, can reduce the skin irritation that is caused by the chemical promoter that contains the skin irritation compositions.

Because the combination of horny layer micropunch and ultrasound application energy adds the extraction speed or the penetrant that use chemical promoter can improve analyte and passes through cuticular transfer rate, therefore, can select concrete carrier and the concrete chemical promoter that uses from prior art in the existing a lot of carrier catalogue, some carriers wherein and chemical promoter were narrated in the above and are incorporated by reference herein.Describe or specifically exemplify in this area existing these chemical compounds in detail and think it is unnecessary.The present invention is not limited to the use of chemical promoter itself, it is believed that, can be used for percutaneous and carry all chemical promoters of medicine in the optics micropunch, to play a role, also can play a role with ultrasonic energy aspect penetrant or the medicine below skin and by skin surface, extracting measurable analyte or carry by skin surface with dyestuff.

Embodiment 16

The mobile ability of synthetic acoustic energy of test and chemical promoter control percutaneous on people's corpse skin sample.In these trials, the thermal release method with embodiment 1 obtains the epidermis film from the separation of people's corpse intact skin.With epidermis film cutting and place between two parts of infiltration box, stratum corneum side is (donor) layer or (receptor) layer down upwards.As U.S. Patent No. 5,445,611 is shown in Figure 2, supports epidermis with the Franz box of repacking.Each Franz box is made up of the upper and lower of clamping with one or several clip.There is a thief hole in lower floor, material promptly from then on the hole add or take down.When upper and lower layer was clipped together, horny layer just was clipped between them.The upper strata of each Franz box makes into to make ultrasonic vibrator to be placed on digression matter tunic 1cm with interior position.With methylene blue as indicator molecules to estimate cuticular infiltration.Process of at every turn being tested with the magnetic tape format that has time mark with video cameras and video tape recorder (not shown) and result's visual record.In addition, take off sample, with seeing through cuticular dyestuff in the absorption spectrometer quantitative assay test.The chemical promoter that is suitable for can be the chemical compound of aforesaid all kinds of solvents and/or interference cell coating.Concrete used promoter is: volume ratio is ethanol/glycerin/water/glyceryl monooleate/methyl laurate of 50/30/15/2.5/2.5.Produce and the system of control acoustic energy comprises that programmable 0-30MHz AWG (Arbitrary Waveform Generator) (Stanford Reserach Systems Model DS345), 20 watts of 0-30MHz amplifiers and formant are respectively 15 and two of 25MHz focus supersonic immersion oscillators not.Prepare 6 boxes simultaneously, be used to test from same donor and the horny layer sample that comes.After placing the horny layer sample, its hydration was tested at least in 6 hours again with distilled water.

Embodiment 17

The effect of acoustic energy during no chemical promoter

As described in embodiment 16, will place the Franz box through the epidermis of thermal release, on the epidermis side direction, under the horny layer side direction, unless refer else.Lower floor fills with distilled water, and the upper strata is filled with the concentrated solution of methylene blue in distilled water.

The epidermis of thermal release: the upper strata is filled to apply acoustic energy in one of them box with complete buried oscillator immediately behind the methylene blue.Such arrangement is placed on the offside of skin fold corresponding to oscillator, or acoustic energy is reflected away from the reflecting plate that is placed on same position, is used for " promotion " analyte to flow out from the opposite side of pleat, enters gathering-device.It is 25MHz that acoustic energy is arranged at the nominal running frequency at first, is equivalent to the intensity of 20 volts of peak-peaks (P-P) input waveform.This roughly is equivalent to the average input energy to 1 watt of oscillator, and is same, supposes that by the data of manufacturer the nominal conversion efficiency of this oscillator is 1%, and then exporting acoustic energy is at 0.78cm ²About 0.01 watt of active regional surface, or acoustic intensity is 0.13 watt/cm ²Other 3 contrast boxes do not have acoustic energy and are applied thereto.Close acoustic energy after 5 minutes.Any box during this time the phase all do not observe dye stream through cuticular macroscopic index, show that dyes concentration is lower than about 0.0015% (v/v) in the 2ml receiver media.

3 the same contrast boxes and the tests of 1 test box are proceeded as follows: the intensity of acoustic energy are increased to from a p-p are input as the output that driver that 70 volts, mean power are input as 12 watts can obtain maximum possible, or about 0.13 watt/cm ²Ultrasonic output intensity.Also frequency configuration is become to be scanned up to 10MHz from 30MHz.The scanning per second of this 20MHz carries out 10 times, and promptly the frequency sweep rate is 10Hz.In these input power level, must monitor the acoustic energy oscillator to avoid overheated.One patch thermocouple is put on the body of oscillator, and the opening and closing power supply remains on below 42 ℃ the maximum temperature of oscillator periodically.Apply peak power after about 30 minutes opening in 1 minute with 50% dutycycle, with the naked eye still can not detect horny layer and permeated by methylene blue with closing in 1 minute.

Then cold water jacket is connected on the acoustic energy oscillator, prolongs the time that on the ceiling capacity level, excites.Adopt same 3 contrasts and 1 experiment box, acoustic energy is put on experiment box last 12 hour with peak power.At this moment, the liquid temperature on upper strata only is elevated to 35 ℃, only a little more than about 31 ℃ at body horny layer normal temperature.Apply acoustic energy as mentioned above after 12 hours, none dye stream occurs through cuticular tangible proof in 4 boxes.

Embodiment 18

The effect of acoustic energy during no chemical promoter

The horny layer of perforation: as described in embodiment 16, prepare 6 boxes.The clamp of clamping Franz box levels to exceeding the required degree of common sealing levels, and is tightened to the degree that manually causes perforation, introduces " pin hole " at thermal release epidermis sample.When dye solution was added to the upper strata of each box, the sign of lower floor bled in the hole that has macroscopic dyestuff to form on horny layer at once.Applying acoustic energy has on the box of such " pin hole " in its mesocuticle thorn, observes liquid transfer of pin hole on horny layer and increases sharply.Whether the transfer rate of indicator dye molecule applies directly related with acoustic energy.That is, applying of acoustic energy causes the instant pulse (lag time approximately＜0.1 second) of indicator molecules through the horny layer pin hole.When cutting off acoustic energy, the pulse of indicator molecules stops (cutting off and lag behind＜0.1 second approximately) immediately.Pulse can be repeated by described.

Embodiment 19

The effect of acoustic energy and chemical promoter

Use two kinds of different chemical promoter preparations.Chemical promoter 1 (CE1) is the mixture of ethanol/glycerin/water/glyceryl monooleate/methyl laurate (50/30/15/2.5/2.5 volume ratio).These compositions are commonly considered as safe as drug excipient by FDA, i.e. GRAS.Chemical promoter 2 (CE2) is a kind of experimental preparation, and it is promoting the medicine percutaneous to be proved to be extremely effective on carrying, but it is generally acknowledged that for the application of long-term percutaneous dosing zest is too big.CE2 contains ethanol/glycerin/water/laurone (lauradone)/methyl laurate (volume ratio 50/30/15/2.5/2.5).Laurone is lauryl (dodecyl) ester of 2-Pyrrolidone-5-carboxylic acid (" PCA "), is also referred to as Laurel acidic group PCA.

Except the placement through the epidermis of thermal release is that epidermal area is downward, promptly on the horny layer side direction outside, as described in preceding (embodiment 16), place 6 Franz boxes.Each sample is exposed to spends the night in the distilled water to finish hydration.When beginning to test, in all 6 boxes, replace the distilled water of lower floor with methylene blue.The upper strata is filled with distilled water, box was observed about 30 minutes, confirm that no dyestuff is by to guarantee all not exist in any box the pin hole perforation.When none is found, from 4 boxes, remove the distilled water on upper strata.Other 2 boxes contrast as distilled water.Then the upper strata of two experiment boxes is filled with CE1, other two experiment boxes fill with CE2.

Apply acoustic energy in two CE2 boxes immediately.Use the oscillator of 25MHz, at 0.13 watt/cm of maximum intensity ≈ ²Per 0.1 second from 10MHz to the 30MHz frequency sweep once.After applying acoustic energy 10-15 minute with 50% dutycycle, naked eyes detect flowing of dyestuff.Not detecting dyestuff in other 5 boxes flows.

Apply acoustic energy in two boxes that contain CE1 of same setting then.In 5 minutes, dyestuff begins to occur on the upper strata.Therefore, acoustic energy and chemical promoter improve the label dyestuff together significantly by cuticular percutaneous flow rate and reduce lag time.

Embodiment 20

The effect of acoustic energy and chemical promoter

With two kinds of chemical promoters, the prescription of CE1 and CE2 deducts glycerol, is prepared, and these new preparations are called CE1MG and CE2MG, as above test.Water consumption substitution glycerol remains unchanged other components in proportions.With 3 boxes of Franz box preparation of improvement, make epidermis side through the epidermis of thermal release to the upper strata in two-layer.Then with the hydration 8 hours in distilled water of these samples.After the hydration step, with the distilled water in CE1MG or the CE2MG replacement lower floor, the upper strata is filled with dye solution.Apply acoustic energy each in 3 boxes in succession.

Apply pulse, frequency modulation acoustic energy total time be less than 10 minutes, the permeability of observing the horny layer sample obviously increases.Be exposed to the whole zone of chemical promoter and acoustic energy, the infiltrative change of horny layer is quite even.Do not observe dyestuff by its leap cuticular " pin hole " puncture.By opening or closing acoustic energy and can control the percutaneous flow velocity immediately.Close the as if instant percutaneous flow velocity that reduces of acoustic energy, down to not seeing that dyestuff passes through skin sample active transport; Possible flow velocity is reduced to the degree of passive diffusion.Open acoustic energy again, recover high-caliber flow velocity immediately.It seems that synthetic mode be the increase that the pulsed percutaneous flow velocity of rule is provided with synthetic speed.When acoustic energy places constant frequency, as if come across about 27MHz for the maximum increase of the percutaneous flow velocity of this set.

After all 3 samples obtain equifinality, discharge all liq, with the cuticular both sides of distilled water flushing.Lower floor fills immediately with distilled water, and the upper strata is filled with dye solution again.Box was observed 30 minutes.On the horny layer sample, do not observe the hole, in lower floor, do not detect a large amount of dyestuffs.Visible a small amount of dyestuff may be the dyestuff and the promoter of catching because of the exposure of skin sample before it in the lower floor.After 12 hours, the amount of dye that detects still seldom.

Embodiment 21

The effect of acoustic energy and chemical promoter

The horny layer of perforation: prepare 3 boxes that have through the epidermis sample of thermal release, the epidermis side to as described in embodiment 16 from the upside of same supplier's box.With sample hydration 8 hours, then the distilled water of lower floor is replaced with CE1MG or CE2MG.The upper strata is filled with dye solution.Pin hole on the horny layer makes dyestuff through bleed the box that contains promoter below it of horny layer sample.Apply acoustic energy.Dye molecule advances through the hole immediately fast after applying acoustic energy.As mentioned above, dyestuff the applying directly of hole and acoustic energy and relevant immediately of flowing through fast.

Embodiment 22

The effect of acoustic energy and chemical promoter

Test the ability of the percutaneous of acoustic energy oscillator TDK#NB-58S-01 (TDK Corp.) raising cheaply flow velocity.The peak response that records this oscillator is about 5.4MHz, other local peaks occurs at about 7MHz, 9MHz, 12.4MHz and 16Mhz.

Test this TDK oscillator in 5.4MHz then and improve the ability of percutaneous flow velocity with CE1MG.3 boxes are set, and the epidermis side is to lower floor, with skin sample hydration 8 hours.In lower floor, put into dye solution.Oscillator is placed on the upper strata to be dipped among the CE1MG.Rate of scanning with 5.3-5.6Mhz excites as acoustic energy, and a large amount of dyestuffs move through horny layer in 5 minutes, are detected in the collection well of box.Local pyrexia occurs, oscillator reaches 48 ℃ of temperature.Do not have in the reference examples of acoustic energy using CE1MG, the exposures in 5 minutes that the dyestuff that obtains in the well is less than use acoustic energy are collected in 24 hours be exposed to.

Present embodiment proves, low-cost, low frequency sound energy oscillator and suitable chemical promoter share, and can influence the percutaneous flow velocity significantly.Although high-frequency sound energy can concentrate on horny layer with more energy in theory, when share with chemical promoter, lower frequency modulation acoustic energy can improve the percutaneous flow velocity, makes this technology become useful and be of practical significance.

Embodiment 23

Confirmation through the molecular migration of application on human skin

Carry out repetition with the test of TDK oscillator and CE1MG at about 12.4MHz (the highest particular resonance peak of oscillator) as mentioned above, frequency scanning is swept to 12.8MHz with 2Hz speed from 12.5, and acoustic density is lower than 0.1W/cm ²Under the epidermis side direction of the epidermis of thermal release, dye solution is in lower floor, and accelerator solution and acoustic energy place the upper strata.The a large amount of dyestuffs of 5 intramolecularly enter the collection well through horny layer.Ohm heating of oscillator is starkly lower than the same oscillator in the 5.4MHz operation, causes that the chemical promoter temperature increases to only about 33 ℃.

Even at like this low level of efficiency, be tangible the detection side to the result of gained with CE1MG with from the acoustic energy of TDK oscillator.U.S. Patent No. 5,445, Fig. 3 A of 611 and 3B show from the curve chart in the data of 3 box gained that detect direction detection percutaneous flow velocity.Even on 5 minutes this point, in the chemical promoter in the horny layer outside, also there is the amount of dye that measures easily, show that dyestuff flow to " outside " of skin sample through horny layer from the epidermis side.

Handle the method that is applied to collect and detect analyte in order to optimize acoustic energy or acoustic energy/chemical promoter, need the instrument of test analyte of interest amount from body.A kind of extract with acoustic energy or acoustic energy/chemical promoter obtain a plurality of readings in the analyte process test macro need not cover on the basis of large sample unified and make different skin properties and flow velocity standardization.Get two or more data points in the time that analyte concentration increases in gathering system with respect to the time, but, thereby set up measuring of interval concentration with regard to the definite parameter of describing analyte take-off speed or flow velocity and reaching the relation curve of equilibrated point of application curves fitting process.The general type of this curve one individual and another be constant between individual, parameter change only.In case these parameters are established, obtain the steady state solution (promptly being equivalent to the time) of this function for infinitely great, i.e. separating when balance is set up just can obtain the concentration of the interior analyte of body.Therefore, this method can be in the mensuration of carrying out required precision for all members of colony (not considering the individual variation of dermal osmosis) with the time of same length.

There are several existing detection techniques to be applicable to the application at present.See D.A.Christensen, in 1648Proceedings of Fiber Optic, Medical and Fluorescent Sensors and Applications 223-26 (1992).A kind of method relates to a pair of fiber optics that use is put together in almost parallel mode very near-earth.One of fiber is the light source fiber, and luminous energy is through this fiber conduction.Another root fiber is the detection fibers that is connected in light sensitive diode.When light conducted by the light source fiber, the part of luminous energy (fast subwave) was present in fiber surface, the detected fiber collecting of the part of this luminous energy.The energy conduction of the fast subwave that detection fibers will be caught is given light sensitive diode, and the latter measures this energy.Fiber adhesive is handled to attract and bonding analyte to be measured.When being incorporated into the surface, the analyte molecule (for example is incorporated into the analyte glucose of fixed agglutinin such as concanavalin A or fixed anti-glucose antibody), the amount of coupled fast subwave changes between two fibers, and the amount of the energy that detected fiber is caught and recorded by diode also changes.Record the measurement several times of fast subwave at short notice, but just fast measuring is described the parameter of the profile of equilibrium, thereby can calculate the concentration of analyte in the body.Show that with this system the mobile experimental result (U.S. Patent No. 5,445,661 Fig. 3 A and 3B) that can measure collected enough data to make accurate last reading in 5 minutes in 5 minutes.

In its most basic embodiment, the device that can be used to apply acoustic energy and collect analyte comprises the baffle of a natural or synthetic material, and it is as the bank of chemical promoter (if you are using) with in order to receive analyzed bank from skin surface.This plate or bank are passively or be fixed on the appropriate location of skin surface institute favored area by means of suitable fastened tools (as belt or adhesive tape).

The placement of acoustic energy oscillator is to make plate or bank between skin surface and oscillator, fixes with proper tools.Power supply is connected on the oscillator, with switching device or any other is suitable mechanically activated.Oscillator is carried the acoustic energy of frequency modulation, phase modulation or amplitude modulation after starting as required, to carry chemical promoter by skin surface from bank, collects analyte to bank from skin surface then.After the required fixing or variable time, oscillator is turned off.The plate or the bank that contain analyte of interest now can take off, any conventional chemical analytical method of using with laboratory or analyte is quantitative with portable instrument.Perhaps, the machinery of quantitative analysis analyte can be contained in and be used to collect in the device of analyte, as the integral part of device or as an adnexa.U.S. Patent No. 5,458,140 have described the device that detects analyte, and this patent is drawn at this and to be reference.

Embodiment 24

Can reach by using Enzymology method in the another kind of method that after the skin surface of perforation is collected sample, detects analyte (as glucose) as mentioned above.The Enzymology method that glucose in the multiple measurement biological sample is arranged.A method relates to the glucose in the glucoseoxidase oxidation sample to produce gluconolactone and hydrogen peroxide.In the presence of colourless chromogen, hydrogen peroxide is converted into water and coloured product then with peroxidase.

Glucoseoxidase

Glucose-------------→ gluconolactone+H ₂O ₂

2H ₂O ₂+ chromogen--------→ H ₂The coloured product of O+

The intensity of coloured product will be proportional with the amount of glucose in the liquid.Color can be measured with the absorption of routine or the method for reflection.Calibrate with known concentration of glucose, i.e. concentration of glucose in the definite analyte of collecting of the amount of available color.Determine its relation by testing, can calculate the concentration of glucose in experimenter's blood.This knowledge can be used in the same way with using the knowledge from finger puncture blood-glucose test gained.The result can obtain in 5-10 minute.

Embodiment 25

Any system that adopts visual display or read concentration of glucose indicate the medicine that needs throwing and insulin or other suitable can for diagnostician or patient.Maybe need to continue to monitor and almost need to take simultaneously under other situation of corrective measure in Intensive Care Therapy, display can be connected with the suitable triggering insulin or the proper signal device of other medicines administration.For example, the insulin pump of implanting abdominal cavity or other body cavity is arranged, it can be activated by exogenous or endogenous stimulus.Perhaps, utilize horny layer micropunch and other technology that improves the percutaneous flow velocity of the present invention, can install the system that percutaneous is carried insulin, its flow speed control is subjected to modulate from the signal of glucose impression system.By this way, can obtain the system of biomedical fully control, it is not only monitored and/or the diagnostic medicine demand, and corrective measure is provided simultaneously.

Ejusdem generis biomedical control system also can be used under other situation, as keeps electrolyte balance or give analgesics according to the analyte parameter (as prostaglandin) that records.

Embodiment 26

When running into another and have medium of different nature with, ultrasound wave, can reflect, reflect and absorb (D.Bommannan et al., 9 with the acoustic phase that can hear Pharm.Res.559 (1992)).Available reflecting mirror or lens focus on or control the structural distribution that it is being studied with acoustic energy.For a lot of positions on the human body, can find that a live body supports this system.For example, ear-lobe is position easily, and available reflecting mirror or lens are helped control of azimuth (passing through horny layer as " promotion " analyte or penetrant), and its effect is similar with intensity of acoustic wave with the change audio frequency.

Embodiment 27

Available a plurality of acoustic energy oscillator is optionally controlled by cuticular percutaneous and is flowed in the body or from the effusive flow direction of body.A skin as ear-lobe, can allow oscillator to place its either side.Oscillator optionally excites or excites with phased approach, flows with the percutaneous that promotes required direction.Can set up the array or the acoustics loop of oscillator, utilize the notion of phased array to carry out (be similar to radar and microwave communication system are invented), make the acoustic energy aim at and focus on the zone of being studied.

Embodiment 28

In the present embodiment, except using excimer laser (as the EMG/200 type of Lambda Physik earlier by U.S. Patent No. 4,775,361 described methods; 193nm wavelength, 14ns pulse width) handle epidermis sample through thermal release with the ablation horny layer outside, carry out according to the method for embodiment 19, this United States Patent (USP) draws at this and is reference.

Embodiment 29

In the present embodiment, except earlier with iodate 1,1 '-diethyl-4,4 '-carbocyanine (Aldrich, λ=703nm) processing is used TOLD9150 type laser diode (Toshiba AmericaElectronic then through the epidermis sample of thermal release, 30mW is in 690nm) conveying total amount 70mJ/cm ²The laser of/50ms to the sample of handling through dyestuff with the ablation horny layer outside, carry out according to the method for embodiment 19.

Embodiment 30

In the present embodiment, be indocyanine green (Sigma cat.No.I-2633 except dyestuff; λ=775nm), laser instrument are outside the Diolite 800-50 (LiCONiX, 50nW is in 780nm), carry out according to the method for embodiment 29.

Embodiment 31

In the present embodiment, except dyestuff is that methylene blue, laser instrument are outside the SDL-8630 (SDL Inc., 500mW is in 670nm), carry out according to the method for embodiment 29.

Embodiment 32

In the present embodiment, except dyestuff is included in the solution that contains penetration enhancer (as CE1), carry out according to the method for embodiment 29.

Embodiment 33

In the present embodiment, ultrasonicly dyestuff and the solution that contains promoter are transported to the horny layer helping, are undertaken by the method for embodiment 29 except being exposed to.

Embodiment 34

In the present embodiment, except light-pulse generator be in the emission of the wide scope of 400-1100nm but in system, be equipped with bandpass filter with export-restriction to the short arc light modulation of about 650-700nm wavelength zone, undertaken by the method for embodiment 31.

Embodiment 35

In the present embodiment, except through the epidermis sample of thermal release earlier with corrected miniature perverse blood pin (BectonDickinson) puncture, generation micropore on horny layer and not reaching outside the tissue below it is undertaken by the method for embodiment 19.

Embodiment 36

In the present embodiment, use 70-480mJ/cm earlier except epidermis sample through thermal release ²The acoustic energy of the focusing of/50ms scope is handled with outside the ablation horny layer, is undertaken by the method for embodiment 19.

Embodiment 37

In the present embodiment, be not more than the micropore of about 100 μ m to form diameter, undertaken by embodiment 19 except carrying out the hydraulic pressure puncture with high-pressure liquid jet earlier.

Embodiment 38

In the present embodiment, except earlier the horny layer puncture being not more than the micropore of about 100 μ m to form diameter, undertaken by embodiment 19 with short electric pulse.

Embodiment 39

Acoustics causes stream effect (Aconotic Streaming)

To describe acoustic energy below is transported to therapeutant in the body and/or body fluid is gathered in the crops the outside in body and store new mechanism and application the storehouse at the micropore that forms on horny layer.Another aspect of the present invention be utilize acoustic energy to around the epidermis of application on human skin and the intact cell in the corium and the fluid between the cell produce acoustics and cause the stream effect.Many document descriptions are arranged, acoustics cause the stream effect be the interactional a kind of pattern of acoustic energy and liquid medium (Nyborg, physical Acoustics Principles and Methods, B p.265-331.Vol.II-Part, AcademicPress.1965).The theory analysis first that acoustics causes flow phenomenon is to be made by Rayleigh (1884,1945).In the extensive discussions to this problem, Longuet Higgins (1953-1960) has drawn near the two-dimentional mobile result who produces applicable to any vibration column face.Nyborg (1958) has developed the three-dimensional of arbitrary surfaces approximate.As Fairbanks et al., 1975 Uitrasonics Symposium Proceedings, IEEE Cat.#75, CHO994-45U is described, acoustic energy and acoustics cause flow phenomenon has very big use to promoting liquid to flow through porous media, show that measurable flow velocity increases up to 50 times, this may be passive or the gradient of only exerting pressure.

All effort that utilize hyperacoustic percutaneous to carry or extract in the past all concentrate on the acoustic energy and the interactional method of skin histology that make horny layer be easy to permeate of being intended to.Related interactional definite mode is assumed to be bar none because the part of horny layer temperature raises, and the fusion (Srinivasan et al.) in the lipid district of the intercellular substance between horn cell that is caused.Other researcheres propose horny layer and form little cave and/or structure and tear and opened passage, and liquid can more easily flow through this passage.In general, the design of the sound system of raising percutaneous flow velocity is based on so early stage understanding, promptly use existing design and be used for the experimenter is produced the ultrasonic therapeutic device of " dark heating " effect, intend importing the gel of intravital medicine or the topical application of aqueous preparation in conjunction with containing, can enter intravital flow velocity to medicine and produce and quantitatively to increase.Disclosed in this manual on the ins and outs that produce the micropore method on this barrier layer, use the mode of acoustic energy to compare with the sound swimming notion of classics definition, can think new fully and different ideas.

Based on United States Patent (USP) 5,458,140 and 5,445,611 described experiments are found, when promptly the horny layer (SC) in the used Franz box of in vitro study is gone up existence or formed duck eye, apply the liquid storage storehouse of the ultrasonic vibrator of suitable driving in any side of SC sample through boring a hole, can produce " acoustics causes stream " phenomenon, wherein liquid greatly the flow velocity pump pressure by this through the perforation film.

With this description shown in the method for the little perforating action that on the skin of living body horny layer, produces control, now practically the liquid stream mode of discussion sound/liquid phase mutual effect liquid suck or the sucking-off body on application.For example, clinical research shows, at 400 μ m ²The micropore of a series of 4 the diameter 80 μ m of last formation applies slight suction (10-12 inch Hg) to this zone then, gets final product the interstitial fluid (ISF) of the on average about 1 μ l of sucking-off, leaves body and is collected in the external vessel.Add the small-sized sound of low-power oscillator in this system, be configured to its inside concentric circular pressure wave of assembling of the tissue of 2-6mm generation around site of perforation quick-actingly, verified, this ISF flow velocity can be enhanced about 50%.

Directly do not absorb (as requiring to produce heat) if do not want on skin histology the acoustic energy that produces certain form, can measure the frequency of skin histology, promptly in the extremely low frequency district of 1kHz to 500kHz the acoustic energy of its substantially transparent.Use the microscopic examination in vivo test, wherein experimenter's skin is by micropunch, and ISF is drawn body and is collected to skin surface, causes the stream effect even also can be observed tangible acoustics in some low-limit frequency of being tested.Make oscillator excited target, show that prominent acoustics causes the macroscopic sign of fluxion amount, because when the ISF whirling motion, little particulate matter is motion together thereupon.Typical motion amplitude can be described below: for the circular ISF of skin surface diameter 3mm converged thing, a visible macroscopic granule had changeed 3 complete circumference each second approximately.This is equivalent to the fluid line speed of second greater than 2.5mm/.Use less than 100mW/cm ²These whole effects of the acoustic energy levels susceptible of proof that enters tissue.

People can easily observe the activity of the upper surface and the top liquid thereof of skin, but the dynamic change that evaluating skin organized layer inside takes place in response to this coupling when acoustic energy is coupled to these organization internals is suitable difficulty.People can suppose, if can so easily produce from the teeth outwards big like this fluid line speed (as＞2.5mm/S), the liquid stream that then is present in dermal tissue intercellular channel alive also should be seen the increase in response to this acoustic energy input.Recently, to applying low frequency acoustic energy zone in the circumference and the ISF that collects through micropore has carried out quantitative measurement around the site of perforation.In this experiment, with identical instrument but load onto oscillator, only replace technology based on the collection ISF of light suction (Hg during 10-12).In a series of 10 acquisition times of two minutes, 55 add acoustic energy with suction only with suction, observe the ISF high approximately 50% that starts sound source and collect in the same time.These data are seen Figure 30.Such ISF flow velocity increases, and is not report any feeling of causing because of acoustic energy the experimenter to realize under the enhanced situation.The used instrument of this test is illustrated in Figure 31-33.The transducer assemblies of Figure 31-33 comprises the thick cyclinder of a piezoelectric, the about 8mm of internal diameter, wall thickness 4mm.With this cylinder polarization, make that the thickness of cylindrical wall is expanded or shunk with the polarity of electric field when applying electric field between the metalized surface of external diameter and internal diameter.In fact, such structure can obtain a kind of device, and this device pushes the tissue that is inhaled in the centre hole rapidly, and the inside radially acoustics of liquid generation that is present in these tissues is caused the stream effect.This inside acoustics causes the micropunch position that the stream effect can be taken more ISF to hole central authorities, leaves body by collected outside at this.

Set up and tested shown in Figure 34 A-B and similarly installed, produce similar PRELIMINARY RESULTS.In Figure 34 A-B sketch map, with salt lake city, Utah State Zevex, the ultrasonic vibrator that Inc. makes is improved, and is added with the spatulate extension on ultrasonic sound collector.The hole that 4mm is arranged at 0.5 of this extension thick scraper end.When starting, main motion is the lengthwise movement along the length of scraper, causes quick back and forth movement basically.The hole that 4mm is set causes the physics fluctuation of metallic spatula, cause in this very active, but unordered big displacement.In use, experimenter's skin is picked up, and enters this hole, in similar mode shown in Figure 33 acoustic energy is imported skin then.

The new aspect of hyperacoustic this new application is following elemental range:

1. no longer need to resemble Langer, Kost, the increase SC that like that ultransonic effect concentrated on that Bommannan etc. proposed stops on the permeability of the membrane.

2. can utilize the lower system of frequency, it seldom is absorbed in skin histology, causes flow phenomenon but still can produce required liquid in the intercellular channel that contains between the epidermis cell of interstitial fluid.

3. ultrasonic and tissue and wherein the interaction mode of liquid be so-called " causing stream " pattern, in the acoustics document, be considered to and the different unique mode of classical oscillating mode interaction that can tear cell membrane, the passive diffusion process of promotion.

Confirmed geometry, frequency, the power of ultrasonic transducer and put on its modulation optimization, can make through perforation skin part liquid stream to be significantly increased.The optimization of these parameters is designed to liquid flow is arranged in investigation in the environment of this microscope yardstick non-linear relation.With the frequency that is lower than 200kHz, can be observed big liquidation effect, and without any the heating that can detect or other bad tissue interaction.It is very low to produce the required acoustic energy levels of these effects that can measure, and average power level is typically at 100mW/cm ²Under.

Therefore, the foregoing description is some representational systems, they can be used to ultrasonic or ultrasonic+chemical promoter is used for collecting and the quantitative assay analyte for diagnosis and percutaneous conveying penetrant.The present invention is directed to following discovery, promptly after cutin perforation, by ultransonic appropriate use, when particularly using with chemical promoter, but not damaged or damage minimum ground percutaneous and measure analyte or conveying penetrant.But the present invention is not restricted to these specific descriptions.Detect or extract some analyte or carry penetrant by horny layer, a lot of puncturing techniques and accelerator systems are arranged, wherein may some be better than other effectiveness.Yet, by the ultimate principle that this paper proposed, those skilled in the art can easily carry out a certain amount of experiment with obtain best perforation, promoter or apply hyperacoustic the most in good time between, intensity and frequency, and the modulation of the frequency of ultrasonic that is applied, amplitude and phase place.Therefore, scope of the present invention only is subjected to the restriction of following claim and function equivalent scope thereof.

Claims (54)

1. the equipment of analyte concentration in the detection individuality is used to promote the permeability of a horny layer of surface selection area to analyte, comprising:

(a) thus with the perforation of the horny layer of institute's favored area, on horny layer, form micropore and do not cause to its down the major injury of covering weave reduce the device of the barrier that horny layer withdraws from analyte;

(b) device of a selected amount of analyte of collection; With

(c) device of the analyte of quantitative assay collection;

It is characterized in that, described device with horny layer perforation be selected from following any: (a) general≤about 1000 μ m ²Thereby remove the cuticular device of institute's favored area on the evaporation point that the horny layer of the selection area of area contacts the horny layer of ablating with a thermal source, make knot of tissue Heshui in the selection area and other temperature that can vaporize material be elevated to described water and other material of can vaporizing; (b) use miniature perverse blood pin once calibration puncture described horny layer, form the device of the micropore of diameter≤about 1000 μ m; (c) with high pressure liquid stream horny layer is carried out the device of hydraulic pressure puncture with the micropore of formation diameter≤about 1000 μ m.

2. equipment as claimed in claim 1, it is characterized in that, it comprises also and applies the acoustic energy of frequency in about 5kHz to 100MHz scope in the described device of selection area through perforation that wherein said acoustic energy is to modulate by a kind of mode that is selected from frequency modulation, amplitude modulation, phase modulation and combination thereof.

3. equipment as claimed in claim 2 is characterized in that, it also comprises makes individual institute's favored area contact and apply the device of acoustic energy with the extraction of further promotion analyte with chemical promoter.

4. equipment as claimed in claim 1, thus wherein said punching machine is to contact, make knot of tissue Heshui and other temperature that can vaporize material to be elevated to the cuticular device of removing institute's favored area on the evaporation point of described water and other material of can vaporizing with thermal source the zone that described cuticular width is not more than about 1000 μ m.

5. equipment as claimed in claim 4, it shows that with the paired pulses light emitted scope of effective dose the dyestuff of absorption is handled described selection area by force, and will be on described dyestuff from a series of pulse concentrations of light-pulse generator output, described dyestuff is heated to be enough to conduct heat to described horny layer, knot of tissue Heshui and other temperature that can vaporize material of institute's favored area is elevated on the evaporation point of water and other material of can vaporizing, wherein said dyestuff plays the thermal source effect.

6. equipment as claimed in claim 5, wherein said light-pulse generator is with the wavelength emission of 100-12000nm.

7. equipment as claimed in claim 6, wherein said light-pulse generator are at about 630-1550nm emitted laser diode.

8. equipment as claimed in claim 6, wherein said light-pulse generator is with the laser diode pumping, at the optical parametric oscillator of about 700-3000nm emission.

9. equipment as claimed in claim 5, wherein said light-pulse generator is selected from arc light, electric filament lamp and light emitting diode.

10. equipment as claimed in claim 5 is characterized in that, it also is provided with the sensor-based system of determining when cuticular barrier has been overcome.

11. equipment as claimed in claim 10, wherein said sensor-based system comprises that reception focuses on light collecting device on the photodiode from the light of described selection area reflection with described reflected light, receive the light and the photodiode that sends a signal on the controller of described focusing, wherein said signal refers to described catoptrical character, and is connected in described photodiode and described light-pulse generator to receive described signal and close the controller of described light-pulse generator after receiving the signal of preliminary election.

12. equipment as claimed in claim 5 is characterized in that, it also has cooling described cuticular selection area and proximate skin tissue, makes institute's favored area and proximate skin be organized in the chiller that is in before the perforation under selected pre-cooling, the limit.

13. equipment as claimed in claim 12, wherein, described chiller comprises a Peltier device.

14. equipment as claimed in claim 5, wherein, described ablation causes interstitial fluid to flow out, and described analyte is to collect in the described interstitial fluid of selected quantity.

15. equipment as claimed in claim 14 is characterized in that, it is after collecting described selected amount of tissue interstitial fluid, and the energy that applies from the effective dose of described light-pulse generator seals described micropore, to cause the interstitial fluid cohesion that is retained in the described micropore.

16. equipment as claimed in claim 14 is characterized in that, it also has the vacuum of applying in cuticular described cuticular selection area through perforation, in order to promote the effusive device of interstitial fluid.

17. equipment as claimed in claim 5 is characterized in that, it uses the unfocused light from described light-pulse generator to shine described selection area at least before with described horny layer perforation, makes the favored area sterilization by described light-struck institute.

18. equipment as claimed in claim 4, it also has the device that institute's favored area is contacted with tinsel, wherein said tinsel plays the thermal source effect, the temperature of institute's favored area is elevated to greater than 100 ℃ from peripheral skin temperature in about 10-50ms, in about 30-50ms, make the temperature of institute's favored area get back to the temperature that is about surrounding skin then, wherein this elevated temperature repeats repeatedly with the circulation of getting back to about peripheral skin temperature, and making is enough to reduce cuticular barrier.

19. equipment as claimed in claim 18 is got back to the device that is about peripheral skin temperature thereby it also has tinsel recalled not contact with horny layer.

20. equipment as claimed in claim 19, it also has between monitoring described tinsel and the individuality by the device of the electrical impedance of described horny layer institute's favored area and proximate skin tissue and advances device wiry, described propulsion plant advanced tinsel when the generation resistance of ablating was reduced thereupon, made in the tinsel heating process to contact with horny layer.

21. equipment as claimed in claim 20, it also has from recalling described device wiry with the horny layer position contacting, wherein said monitoring device can detect the variation that contacts the impedance of being followed with horny layer following table cortex and transmit signal to the described device of recalling to recall tinsel, it is not contacted with horny layer.

22. equipment as claimed in claim 19, wherein, described tinsel heats with the Ohmic heating element.

23. equipment as claimed in claim 19, wherein, described tinsel forms the circuit that comprises high resistance point, and the temperature of described high resistance point is by being modulated modulated electric current by described circuit.

24. equipment as claimed in claim 19, wherein, described tinsel is arranged in the modulated AC magnetic field of magnet exciting coil, makes when passing to alternating current to magnet exciting coil, can produce the eddy current that is enough to by inner ohmic loss heating of metal silk.

25. equipment as claimed in claim 1, wherein, described punching machine be with the miniature perverse blood pin of calibrating puncture described horny layer, form the device that diameter is not more than the micropore of about 1000 μ m.

26. equipment as claimed in claim 1, wherein, described punching machine be with high pressure liquid stream hydraulic pressure puncture described horny layer, form the device that diameter is not more than the micropore of about 1000 μ m.

27. as claim 1,5 or 18 described equipment, wherein, analyte is a glucose.

28. equipment as claimed in claim 27, wherein, glucose quantitatively be to carry out colorimetric determination or by biosensor by glucoseoxidase.

29. one kind is used to promote that the active substance percutaneous flows into the equipment of individual selection area, is used to promote the permeability of the horny layer in a selected zone, surface to described active penetrant, it comprises:

(a) thus with the perforation of the horny layer of institute's favored area, on horny layer, form micropore and do not cause to its down the major injury of covering weave reduce the device of horny layer to the barrier of described active permeate flow;

(b) thus the institute's favored area through perforation will be contacted the device that promotes described penetrant inflow body with the compositions that comprises the described penetrant of effective dose,

It is characterized in that, described device with horny layer perforation be selected from following any: (a) general≤about 1000 μ m ²Thereby remove the cuticular device of institute's favored area on the evaporation point that the horny layer of the selection area of area contacts the horny layer of ablating with a thermal source, make knot of tissue Heshui in the selection area and other temperature that can vaporize material be elevated to described water and other material of can vaporizing; (b) use miniature perverse blood pin once calibration puncture described horny layer, form the device of the micropore of diameter≤about 1000 μ m; (c) with high pressure liquid stream horny layer is carried out the device of hydraulic pressure puncture with the micropore of formation diameter≤about 1000 μ m.

30. equipment as claimed in claim 29, it is characterized in that, thereby it also has and applies acoustic energy and promote described penetrant percutaneous to flow into the device of body in described intensity and the frequency that produces the effect make liquid flow through the regional regular hour of perforation, effectively.

31. equipment as claimed in claim 30, wherein, the frequency of described acoustic energy is about 5kHz to 100MHz, and wherein said acoustic energy is modulated by a kind of mode that is selected from frequency modulation, amplitude modulation, phase modulation and combination thereof.

32. equipment as claimed in claim 31 is characterized in that, institute's favored area that it also has individuality contacts and applies acoustic energy flows into the body of described individuality with the described penetrant of further promotion device with chemical promoter.

33. equipment as claimed in claim 29, wherein, thus described punching machine is a described cuticular described selection area that width is not more than about 1000 μ m contacts, makes selected regional organization bound water and other temperature that can vaporize material to be elevated to the cuticular device of removing institute's favored area on the evaporation point of described water and other material of can vaporizing with a thermal source.

34. equipment as claimed in claim 33, it shows the strong dyestuff treatment of selected zone that absorbs with the paired pulses light emitted scope of effective dose, and will be on described dyestuff from a series of pulse concentrations of described light-pulse generator output, described dyestuff is heated to be enough to conduct heat to described horny layer, selected regional organization bound water and other temperature that can vaporize material are elevated on the evaporation point of described water and other material of can vaporizing, wherein said dyestuff plays the thermal source effect.

35. equipment as claimed in claim 34, wherein, the wavelength of described light-pulse generator emission 10-12000nm.

36. equipment as claimed in claim 35, wherein, described light-pulse generator is the scope emitted laser diode at about 630-1550nm.

37. equipment as claimed in claim 35, wherein, described light-pulse generator is the optical parametric oscillator of launching in about 700-3000nm scope with the laser diode pumping.

38. equipment as claimed in claim 35, wherein, described light-pulse generator is a kind of light source that is selected from arc light, electric filament lamp and light emitting diode.

39. equipment as claimed in claim 34, it also has sensor-based system, in order to determine when cuticular barrier is excluded.

40. equipment as claimed in claim 39, wherein, sensor-based system comprises light that reception comes from the reflection of described selection area and the light collecting device that described reflected light is focused on photodiode, receive the light and the photodiode that sends a signal to controller of described focusing, the described catoptrical character of wherein said signal indication, and be coupled to described photodiode and the controller of described light-pulse generator to receive described signal and behind the signal that receives preliminary election, to cut off described light-pulse generator.

41. equipment as claimed in claim 34, it also has the cooling of described cuticular institute's favored area and proximate skin tissue, makes described zone and proximate skin be organized in chiller under the steady-state condition that is in selected pre-cooling before the puncture.

42. equipment as claimed in claim 41, wherein, described chiller comprises the Peltier device.

43. equipment as claimed in claim 34, it also has the described institute of the unfocused rayed favored area of using at least from described light-pulse generator, the device that uses the favored area sterilization of described light-struck institute before described horny layer perforation.

44. equipment as claimed in claim 33, it also has the device that described selection area is contacted with tinsel, wherein said tinsel plays the thermal source effect, the temperature of institute's favored area is elevated to greater than 100 ℃ from peripheral skin temperature in about 10-50ms, in about 30-50ms, make the temperature of institute's favored area get back to the temperature that is about surrounding skin then, wherein this elevated temperature repeats repeatedly with the circulation of getting back to about peripheral skin temperature, and making is enough to reduce cuticular barrier.

45. equipment as claimed in claim 44 is got back to the device that is about peripheral skin temperature thereby it also has described tinsel recalled not contact with horny layer.

46. equipment as claimed in claim 45, it also has between monitoring described tinsel and the described individuality by the device of the electrical impedance of described horny layer institute's favored area and proximate skin tissue and advances the device of described tinsel position, make when ablation taking place and follow the reduction of resistance, described propulsion plant can advance tinsel, contacts with horny layer when making the tinsel heating.

47. equipment as claimed in claim 46, it also has the device of described tinsel from recalling with the horny layer position contacting, wherein monitoring device can be measured with horny layer following table cortex and contact the impedance variation of being followed and transmit signal to the described device of recalling, with described tinsel from recalling with the horny layer position contacting.

48. equipment as claimed in claim 45, wherein, described tinsel is to heat with the Ohmic heating element.

49. equipment as claimed in claim 45, wherein, described tinsel forms the current loop that comprises high resistance point, and the temperature of described high resistance point is by modulated by described current loop by synthetic electric current.

50. equipment as claimed in claim 45, wherein, described tinsel place can synthetic magnet exciting coil AC magnetic field, can produce the eddy current that is enough to by inner ohmic loss heated wire when making magnet exciting coil pass to alternating current.

51. equipment as claimed in claim 29, wherein, described punching machine be with the miniature perverse blood pin of calibrating puncture described horny layer, form the device that diameter is not more than the micropore of about 1000 μ m.

52. equipment as claimed in claim 29, wherein, described punching machine be with high pressure liquid stream hydraulic pressure puncture described horny layer, form the device that diameter is not more than the micropore of about 1000 μ m.

53. apply the equipment of tattooing on the selection area of individual surface skin, it comprises with lower device:

(a) thus with the perforation of the horny layer of institute's favored area, on horny layer, form micropore and do not cause to its down the major injury of covering weave reduce the device of horny layer to the barrier of active permeate flow; With

(b) will contact, promote described ink to flow into the device of body with the compositions that comprises effective dose tattooing ink through institute's favored area of perforation;

It is characterized in that, described device with horny layer perforation be selected from following any: (a) general≤about 1000 μ m ²Thereby remove the cuticular device of institute's favored area on the evaporation point that the horny layer of the selection area of area contacts the horny layer of ablating with a thermal source, make knot of tissue Heshui in the selection area and other temperature that can vaporize material be elevated to described water and other material of can vaporizing; (b) use miniature perverse blood pin once calibration puncture described horny layer, form the device of the micropore of diameter≤about 1000 μ m; (c) with high pressure liquid stream horny layer is carried out the device of hydraulic pressure puncture with the micropore of formation diameter≤about 1000 μ m.

54. be used to reduce in skin selection area analyte is diffused into the interstitial fluid of described individuality from the blood of individuality the equipment of time delay, it comprises:

(a) thus with the perforation of the horny layer of institute's favored area, on horny layer, form micropore and do not cause to its down the major injury of covering weave reduce the device of the barrier that horny layer withdraws from analyte;

(b) with the refrigerative chiller of described selection area of skin;

(c) see through the device that micropore is collected a selected amount of interstitial fluid, described interstitial fluid contains analyte;

It is characterized in that, described device with horny layer perforation be selected from following any: (a) general≤about 1000 μ m ²Thereby remove the cuticular device of institute's favored area on the evaporation point that the horny layer of the selection area of area contacts the horny layer of ablating with a thermal source, make knot of tissue Heshui in the selection area and other temperature that can vaporize material be elevated to described water and other material of can vaporizing; (b) use miniature perverse blood pin once calibration puncture described horny layer, form the device of the micropore of diameter≤about 1000 μ m; (c) with high pressure liquid stream horny layer is carried out the device of hydraulic pressure puncture with the micropore of formation diameter≤about 1000 μ m.

55. be used to reduce the vaporization of interstitial fluid and the equipment of vapour pressure thereof, it comprises:

(a) thus with the perforation of the horny layer of institute's favored area, on horny layer, form micropore and do not cause to its down the major injury of covering weave reduce the device of the barrier that horny layer withdraws from analyte;

(b) with the refrigerative chiller of described selection area of skin;

(c) see through the device that micropore is collected a selected amount of interstitial fluid;

It is characterized in that, described device with horny layer perforation be selected from following any: (a) general≤about 1000 μ m ²Thereby remove the cuticular device of institute's favored area on the evaporation point that the horny layer of the selection area of area contacts the horny layer of ablating with a thermal source, make knot of tissue Heshui in the selection area and other temperature that can vaporize material be elevated to described water and other material of can vaporizing; (b) use miniature perverse blood pin once calibration puncture described horny layer, form the device of the micropore of diameter≤about 1000 μ m; (c) with high pressure liquid stream horny layer is carried out the device of hydraulic pressure puncture with the micropore of formation diameter≤about 1000 μ m.

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