CN1867337A - Methods for intradermal delivery of therapeutics agents - Google Patents

Abstract

The present invention relates to methods and devices for delivering one or more biologically active agents, particularly therapeutic agents, the intradermal compartment of a subject's skin. The present invention provides an improved method of delivery of biologically active agents, such as therapeutic agents, through lymphatic vasculature accessed by intradermal delivery. Therapeutic agents to be delivered in accordance with the present invention include, but are not limited ton antineoplastic agents, chemotherapeutic agents, antibodies, antibiotics, anti-angiogenesis agents, anti-inflammatory agents, and immunotherapeutic agents. Therapeutic agents delivered in accordance with the present invention have improved bioavailability, including improved systemic distribution and improved delivery to particular tissues. Therapeutic agents, delivered in accordance with the methods of the invention have an improved clinical utility and therapeutic efficacy relative to other drug delivery methods, including intraperitoneal, intramuscular and subcutaneous delivery. The methods of the present invention provide benefits and improvements over convention drug delivery methods including dose sparing, increased drug efficacy, reduced side effects, reduced metastatic potential and prolonged survival.

Description

The method that is used for intradermal delivery of therapeutics agents

The application requires the U.S. Provisional Application No.60/550 of submission on March 3rd, 2004, the U.S. Provisional Application No.60/497 that on August 26th, 197 and 2003 submitted to, and 702 priority, and with they complete being collected herein by reference.

1. invention field

The present invention relates to be used for one or more biologic activity agent (particularly therapeutic agent) are delivered to the method and apparatus in intradermal (intradermal) zone (compartment) of experimenter's skin.The invention provides by intradermal delivery biologic activity agent (such as therapeutic agent) is delivered to improving one's methods of lymph vascular system.Can include but not limited to antitumor agent, chemotherapeutant, antibody, antibiotic, antiangiogenic agent, antiinflammatory and immunotherapeutic agent according to the therapeutic agent that the present invention delivers.The therapeutic agent of delivering according to the present invention has the bioavailability of improvement, comprises the whole body distribution of improvement and the particular organization's delivery that improves.With respect to other medicines delivering method (comprising intraperitoneal, intramuscular and subcutaneous delivery), have the clinical efficacy and the therapeutic effect of improvement according to the therapeutic agent of the inventive method delivery.Method of the present invention provides benefit and the improvement that is better than the conventional medicine delivering method, comprises saving dosage, improve effect of drugs, reduce side effect, reduce metastatic potential and prolonging survival.

2. background of invention

People have recognized the efficient and safe importance of using medicament (such as therapeutic agent) and medicine for a long time.Recently shown especially aspect the bioavailability of macromole (such as the protein of being produced by biotechnological industries) being guaranteed appropriateness and the reproducible absorption difficulty (Cleland etc., 2001, Curr.Opin.Biotechnol.12:212-219).The use of conventional needle provides a kind of method of humans and animals being delivered medicament by the administration of passing skin for a long time.Generally speaking, the harsh conditions relevant with oral delivery have been avoided in injection, and these conditions usually weaken the desired effects of most of biological therapies.Injection can also provide therapeutic effect faster than oral administration.In order to realize reproducible and to deliver based on the injection of pin efficiently, improve the facility of using simultaneously and alleviate uneasiness and/or the pain of patient conventional needle, made considerable effort.In addition, some percutaneous delivery system has been eliminated pin fully, drives (such as iontophoretic current (iontophoreticcurrent) or electroporation or hot piercing or phonophoresis (sonophoresis)) and breaks through horny layer (outermost layer of skin) and pass skin surface and deliver medicament and rely on simple hydrophobic absorption, chemical mediator or external force.Yet these delivery systems generally can not reproduciblely cross skin barrier or medicament is delivered to designated depth below the skin surface.Therefore, clinical effectiveness may be changeable.Thus, think that machinery (such as use pin) breakthrough horny layer uses medicament the most reproducible method is provided for passing skin surface, and provide control and reliability for the location of using medicament.

Be used for below skin surface delivering the method for medicament almost exclusive comprise percutaneous injection or infusion, promptly pass skin medicament be delivered to position below the skin.That percutaneous injection and infusion comprise is subcutaneous, intramuscular or intravenous administration approach, and wherein the most frequently used is intramuscular (IM) and subcutaneous (SC) injection.

Anatomically, the outer surface of health is made up of two kinds of vital tissue layers, the epidermis of outside and following corium, the two has constituted skin together, and (summary is seen " physiology of skin, biochemistry and molecular biology " (physiology, Biochemistry, andMolecular Biology of the SKin), the 2nd edition, L.A.Goldsmith compiles, the Oxford University Press, New York, 1991).Epidermis is subdivided into five layers, and thickness adds up to 75-150 μ m.The below the epidermis is a corium, and it comprises two-layer, and outmost part is called mamillary corium, and deep layer is called reticular corium.Mamillary corium contains a large amount of microcirculation blood and lymphatic plexus.On the contrary, reticular corium contains less cell and vascular, is made up of the collagen and the elastic [connective of densification.Be subcutaneous tissue below epidermis and the corium, be also referred to as hypodermis (hypodermis), it is made of connective tissue and fatty tissue.Muscular tissue be positioned at hypodermic below.

As mentioned above, subcutaneous tissue and muscular tissue all usually are used as the medicine-feeding part of medicament (comprising therapeutic agent).Yet seldom as the medicine-feeding part of medicament, this may be that (to small part) is because be difficult to the pin accurate in locating in the intradermal zone to corium.In addition, distribute, yet do not recognize the vascular that can utilize this height to distribute so far as yet and the absorption characteristic that the medicament of institute's administration obtained improve to some extent than subcutaneous administration although known corium (particularly mamillary corium) has the vascular of height.

Conventional a kind of medication of having used below skin surface and having entered the intradermal zone in awns figure (Mantoux) tuberculin test.In this program, use 27 or No. 30 pins to enter skin surface and inject the protein derivatives (Flynn etc., 1994, Chest 106:1463-5) of purification with low-angle.Yet, inject localized to a certain degree uncertainty and may cause some false negative test results.In addition, at the injection site initiating response, this test involves local injection, and Mang Tufa does not cause intradermal injection in whole body administration medicament.

The whole body administration of so-called " intradermal " injection has been reported by some groups.In such portion report, carried out comparative study (Autret etc., 1991, Therapie 46:5-8) about subcutaneous and so-called " intradermal " injection.The medicament of test usefulness is a calcitonin, a kind of protein of molecular weight about 3600.Although claiming medicine is the intradermal injection, yet the 4mm pin is used in injection, and be advanced into the bottom with 60 degree angles.This will cause injection to be positioned at the degree of depth of about 3.5mm, enter the bottom of reticular corium, perhaps enter subcutaneous tissue, but not enter the mamillary corium of the vascular that gathers.In fact,, can expect that so medicament will slowly absorb or is diffused into subcutaneous area in the less reticular corium of vascular, and cause on function with subcutaneous administration and absorb identical if this group has been expelled to the bottom of reticular corium but not has entered subcutaneous tissue.This real or functional subcutaneous administration will be explained in the report between subcutaneous administration and the so-called intradermal administration reason in the concentration of the time that reaches maximal plasma concentration, each minute and area under curve shortage difference aspect these.

Similarly, Bressolle etc. by so-called " intradermal " injection use No. 4 pin administration ceftazidime sodium (sodium ceftazidime) (Bressolle etc., 1993, J.Pharm.Sci.82:1175-1178).This will cause the injection in the following 4mm degree of depth of skin surface, produce real or functional subcutaneous injection, although expect that in this case subcutaneous absorption is better because ceftazidime sodium be hydrophilic and molecular weight lower.

The report of another group is about so-called intradermal drug delivery device (U.S. Patent number 5,007,501).Point out that injection is at a slow speed, and the injection site is intended that some zones of below the epidermis, i.e. interface between epidermis and the corium or corium or hypodermic inside.Yet this piece list of references is not instructed the administration of hint selectivity corium, does not hint the issuable any possible pharmacokinetics advantage of this selectivity administration yet.

Thus, still continue to need efficiently and the method and apparatus of the administration medicament (especially therapeutic agent) of safety.

3. summary of the invention

The invention provides the method that is used for one or more biologic activity agent (preferred therapeutic agents) of percutaneous drug delivery of experimenter, wherein the biologic activity agent is delivered to the intradermal zone of experimenter's skin.In a preferred embodiment, can include but not limited to antitumor agent, chemotherapeutant, antibody, antibiotic, antiangiogenic agent, antiinflammatory and immunotherapeutic agent according to the therapeutic agent that the present invention delivers.

The present invention's part is based on inventor's following discovery, and when soon medicament was delivered to the intradermal zone, they were delivered to the system of regional nodes rapidly, and whole body distributes, and is dispensed to tissue more deeply.Can directly arrive vein and two kinds of networks of lymph of corium according to intradermal delivery of therapeutics agents of the present invention, and unique systemic drug kinetic results is provided.By arriving these networks, just can realize treating benefit including but not limited to improve clinical efficacy and therapeutic effect.Particularly, the inventor finds, therapeutic agent is administered to the intradermal zone causes therapeutic effect to be improved with respect to other medicines delivering method (comprising the intraperitoneal delivery).

In a specific embodiment, the invention provides and be used for improving one's methods of delivery of therapeutics agents, wherein said therapeutic agent includes but not limited to antitumor agent, chemotherapeutant, antibody, antibiotic, antiangiogenic agent, antiinflammatory, immunotherapeutic agent and antiviral agent, and clinical efficacy and therapeutic effect are improved.

Pass through vein and two kinds of network delivery of lymph of corium rapidly according to the biologic activity agent (such as therapeutic agent) of the inventive method administration.The therapeutic agent of delivering according to the inventive method is at the intradermal area deposition, and is distributed to regional nodes's tissue of medicine-feeding part with high bioavailability, and the lymph of broadcasting by propagation is more delivered and entered systemic circulation subsequently.This therapeutic agent delivering method utilizes in the disease of two kinds of networks particularly useful in treatment, include but not limited to cancer, tumor growth and transfer, viral infection, bacterial infection, parasitic infection, immune disorders and metabolic disorder.

Can directly arrive vein and two kinds of networks of lymph of corium according to intradermal delivery of therapeutics agents of the present invention, and unique systemic drug kinetic results is provided.By arriving near these networks of target (such as tumor), just may realize treating benefit.The therapeutic agent of intradermal administration can also arrive immunocyte by the mediation of lymph network.In addition, intradermal delivery of therapeutics agents will cause whole body to distribute, and provide the position of arrival wide dispersion and the added advantage of organ in the mode similar to many current therapy.

The invention provides and improve improving one's methods of the biologic activity bioavailability of agent in particular organization, include but not limited to skin histology, lymphoid tissue (for example lymph node), mucosal tissue, germinal tissue, cervical tissue (cervical tissue), vagina tissue and form and the health any part of carrying out specific function is an organ, include but not limited to lung, spleen, colon, thymus by dissimilar tissues.In some embodiment, tissue comprises any tissue that maybe can arrive environment with environmental interaction, for example skin, mucosal tissue.Other tissue that the present invention is contained includes but not limited to hemolymph sample system; Lymphoid tissue (for example relevant lymphoid tissue of epithelium and mucosa-associated lymphoid tissue or MALT (MALT can further be divided into the lymphoid tissue (D-MALT) of organized mucosa-associated lymphoid tissue (O-MALT) and disperse); Elementary lymphoid tissue (for example thymus and bone marrow); Secondary lymphoid tissue (for example lymph node, spleen, digestion, breathing and urogenital tissue).Those skilled in the art will understand, and secondary MALT comprises gut associated lymphoid tissue (GALT); BALT (BALT); And urogenital system.MALT specifically comprises the conjunctiva of various joint knots, skin and eye in peyer's patches in lymph node, spleen, tissue such as the jaw (palentine) relevant with epithelial surface and nasopharynx tonsil, the small intestinal (Peyer ' spatches) and breathing and the genitourinary system.O-MALT comprises pharynx week lymph sample ring, the esophagus joint knot of tonsil (jaw, tongue, nasopharynx and pipe) and runs through the similar lymphoid tissue that whole digestive tract is scattered to anal canal by duodenum.

Except other benefit, provide following benefit according to intradermal delivery biologic activity of the present invention agent, promptly absorb rapidly and enter regional nodes, institute's medicament of delivering is to the targeting and the deposition raising of particular organization, and whole body and tissue biological's availability raising.These benefits are particularly useful such as therapeutic agents such as antitumor agent, antibody and antibiotic for delivering.Intradermal delivery medicament according to the inventive method is deposited on medicament in intradermal and lymph zone and the deep tissue, causes medicament rapidly and in biologically focusing on these zones and tissue significantly.

With the direct targeting lymph of various therapeutic agents, realized useful therapeutic effect by intradermal delivery, comprised and save dosage, improve effect of drugs, reduce side effect, reduce metastatic potential and prolong life cycle.Since delivering method of the present invention can make therapeutic agent not only whole body but also local deposits, the invention provides so and be used for the treatment of improving one's methods of disease.Therefore, the deposition by improving institute's delivery of therapeutics agents, tissue biological's availability, begin effect (onset) and removing faster faster, the invention provides and be used for the treatment of improving one's methods of disease (such as cancer).The invention provides in order to treat the method for disease (particularly cancer) at least a therapeutic agent of administration, comprise with controllable rate, volume and pressure medicament is administered to the intradermal zone of experimenter's skin, make medicament be deposited in the ID zone and and absorb by the lymph vascular system.

Deposition by improving institute's delivery of therapeutics agents, tissue biological's availability, begin effect and removing faster faster, the present invention also provides and has been used for the treatment of improving one's methods of the disease that is positioned at given body tissue and organ, such as the infection of those tissues and organ, for example respiratory tract infection.The invention provides in order to treat the method for disease (particularly infecting) at least a therapeutic agent of administration, comprise with controllable rate, volume and pressure medicament is delivered to the intradermal zone of experimenter's skin, make medicament be deposited in the intradermal zone and and absorb by the lymph vascular system.

When being used for this paper, be delivered in the intradermal zone or intradermal delivery means the biologic activity agent is administered into mode in the corium, the mamillary corium that makes medicament be easy to arrive to be rich in vascular, and absorb rapidly in blood capillary and/or the lymphatic vessel and to become the general biology available.This can be achieved like this, and being about to medicament, to place the upper area of corium be mamillary corium, and perhaps the top of the less reticular corium of vascular makes medicament be easy to be diffused in the mamillary corium.The biologic activity agent below mamillary corium and in reticular corium but enough be higher than controlled delivery in this dermal zone at interface between corium and the subcutaneous tissue should make medicament effectively (outwards) move to (undisturbed) vascular and lymphatic capillaries bed (in mamillary corium), it can absorb by these capillary tubies and enter the body circulation at this, and can not detained (sequestered) by any other skin histology zone in current process.In some embodiment, with the biologic activity agent mainly place at least about 0.3mm, more preferably at least about 0.4mm and most preferably at least about the degree of depth of 0.5mm to being no more than about 2.5mm, will causing the rapid absorption of medicament more preferably no more than the about 2.0mm and the degree of depth that is most preferably not exceeding about 1.7mm.Although do not want to be subject to concrete mechanism of action, the bottom that the biologic activity agent is mainly placed the bigger degree of depth and/or enter reticular corium may cause lymph effective inadequately to the absorption of medicament, because medicament will slowly absorb in the vascular less reticular corium or subcutaneous area.

With deliver the relevant improvement benefit of biologic activity agent according to the inventive method ID and not only can use and realize based on the injecting systems of micro device, can also use other delivery system to realize, inject with pin (needle-less) or without the impact of pin (needle-free) such as making fluid or powder enter the few of ID zone, by the enhanced iontophoresis of micro device (ionotophoresis), and fluid, solid or other dosage form direct deposition in skin.In specific embodiment, the administration of biologic activity agent is by the intradermal zone of pin or sleeve pipe insertion experimenter skin is realized.

3.1 definition

When being used for this paper, " intradermal " refers to the biologic activity agent is administered into mode in the corium, the mamillary corium that makes medicament be easy to arrive to be rich in vascular, and absorb rapidly in blood capillary and/or the lymphatic vessel and to become the general biology available.This can be achieved like this, and being about to medicament, to place the upper area of corium be mamillary corium, and perhaps the top of the less reticular corium of vascular makes medicament be easy to be diffused in the mamillary corium.The biologic activity agent below mamillary corium and in reticular corium but enough be higher than controlled delivery in this dermal zone at interface between corium and the subcutaneous tissue should make medicament effectively (outwards) move to (undisturbed) vascular and lymphatic capillaries bed (in mamillary corium), it can absorb by these capillary tubies and enter the body circulation at this, and can not detained by any other skin histology zone in current process.In some embodiment, with the biologic activity agent mainly place at least about 0.3mm, more preferably at least about 0.4mm and most preferably at least about the degree of depth of 0.5mm to being no more than about 2.5mm, will causing the rapid absorption of medicament more preferably no more than the about 2.0mm and the degree of depth that is most preferably not exceeding about 1.7mm.Although do not want to be subject to concrete mechanism of action, biologic activity reagent mainly placed the bigger degree of depth and/or enter the bottom of reticular corium or absorption that the SC zone will cause lymph effective inadequately.

When being used for this paper, " intradermal delivery " refers to medicament is delivered to the intradermal zone, as Pettis etc. (with they complete being collected herein by reference) described in WO02/02179A1 (PCT/US01/20782) and the U. S. application series number 09/606,909.

When being used for this paper, " subcutaneous delivery " refers to medicament is deposited in the hypodermic layer of experimenter's skin, and the degree of depth is greater than 2.5mm.

When being used for this paper, " pharmacokinetics, pharmacodynamics and bioavailability " as Pettis etc. described in the WO02/02179A1 (PCT/US01/20782, June 29 2000 priority date).

When being used for this paper, " pharmacokinetics of improvement " refers to compare with conventional medication, behind the medicament of administration specified amount, and (T lag time of the bioavailability of raising, shortening _Lag), shorten T _Max, accelerate absorption rate, begin the C that acts on and/or raise faster _Max

When being used for this paper, " bioavailability " refers to arrive in the administration medicament of prescribed dose the total amount of blood part.This measures the area under curve in the time collection of illustrative plates with concentration usually.

When being used for this paper, " tissue " refers to carry out together a group or a confluent monolayer cells of function, include but not limited to skin histology, lymphoid tissue (for example lymph node), mucosal tissue, germinal tissue, cervical tissue, vagina tissue and form and the health any part of carrying out specific function is an organ, include but not limited to lung, spleen, colon, thymus by dissimilar tissues.When being used for this paper, tissue comprises and environmental interaction or accessibility any tissue, for example skin, mucosal tissue.

When being used for this paper, " tissue biological's availability " refers to the amount of biological available medicament in particular organization in vivo.This tittle is usually may relate to the active of combination, labelling, detection, conveying, stability, biological action or to can be used for diagnosing and/or but other measurement characteristics for the treatment of is measured.In addition, the definition that is to be understood that " tissue biological's availability " also comprises the amount of the available medicament of particular organization." tissue biological's availability " is included in the amount of the medicament that accumulates in the particular organization of extra fine quality/volume in the amount of the medicament that accumulates in the particular organization of the medicament total amount that accumulates in the particular organization, the amount that is the medicament of passing particular organization, unit mass/volume and each unit of time.Tissue biological's availability is included in the inherent particular organization of body or the tissue set (is gathered such as the tissue that constitutes vascular and/or various organs, organ is promptly by dissimilar organizational compositions and carry out the body part of specific function, and example comprises spleen, thymus, lung, lymph node, the heart and brain) in the amount of available medicament.

When being used for this paper, " lag time (lag time) " shows the medicine medicament and reaches the delay that can measure between the time that maybe can detect blood or blood plasma level.T _MaxThe value representative reaches the time of the maximum haemoconcentration of medicament, and C _MaxValue refers to the maximum haemoconcentration that reaches by specified dosage and medication.Beginning action time is T _Lag, T _MaxAnd C _MaxFunction realize the necessary blood of biology effect (or target tissue) concentration time necessary because all these parameters all influence to reach.Can measure T by the macroscopic examination graphic result _MaxAnd C _Max, and usually can provide enough information for two kinds of methods that compare the administration medicament.Yet, can use mathematical model and/or other means that those skilled in the art will know that by this numerical value of the more accurate mensuration of dynamic analysis.

When being used for this paper, any method that " conventional deliver " refers to be used to deliver any material, they have or think and have similar or lower to subcutaneous delivery, improved biodynamics and vitodynamics.That conventional delivery can comprise is subcutaneous, iontophoresis and Intradermal delivering method, such as Gross at US5, described in 800,420.

When being used for this paper, term " disease " and " disease " are used interchangeably, and refer to experimenter's the patient's condition.Disease comprise the function of health (system or organ) any interruption, stop or disorderly.

When being used for this paper, term " cancer " refers to by vegetation or tumor unusual, that uncontrolled cell growth causes.When being used for this paper, cancer clearly comprises leukemia and lymphoma.Term " cancer " refers to involve the disease of such cell, they might be transferred to than the distal part position and show the phenotypic characteristic different with non-cancerous cell, for example form colony or form tubulose network or net sample substrate in three dimensional matrix (such as soft agar) in three-dimensional substrates film or extracellular matrix prepared product.Non-cancerous cell does not form colony in soft agar, and forms unique ball spline structure in three-dimensional substrates film or extracellular matrix prepared product.Cancerous cell has obtained the functional capabilities of a stack features in their evolution, although be by different mechanism.These abilities comprise evades that apoptosis, growth signals are self-sufficient, the antagonism growth signals is insensitive, invade the potentiality of tissue/transfer, infinite multiplication and lasting blood vessel generation.Term " cancerous cell " intention contains cancerate preceding and pernicious two kinds of cancerous cell.In some embodiment, cancer refers to keep the benign tumor that localizes.In other embodiments, cancer refers to invade and destroy contiguous body structure and is transmitted to malignant tumor than the distal part position.In also having some embodiments, cancer is relevant with special cancer antigen.

When being used for this paper, term " experimenter " and " patient " are used interchangeably.When being used for this paper, experimenter's preferred mammal, such as non-human primate (for example cattle, pig, horse, cat, Canis familiaris L., rat etc.) and primates (for example monkey and people), optimum is chosen.

When being used for this paper, " treatment effective dose " refers to be enough to treat or the amount of control disease or treatment of conditions agent.The treatment effective dose can refer to be enough to postpone the outbreak of disease or it is reduced to the amount of minimum therapeutic agent, for example postpones the propagation of cancer or it is reduced to minimum.The treatment effective dose can also refer to provide the amount of the therapeutic agent of therapeutic benefit in treatment of diseases or control.In addition, the treatment effective dose refers to separately or unites other treatment provides the therapeutic agent of therapeutic benefit in treatment of diseases or control amount for therapeutic agent of the present invention.

When being used for this paper, term " preventive " refers to can be used for preventing any medicament of disease or its recurrence or propagation.

When being used for this paper, " prevention effective dose " can refer to be enough to prevent the amount of the preventive that excess proliferative disease (particularly cancer) takes place, recurs or propagate in the patient, include but not limited to easily suffer from the patient of excess proliferative disease, for example the easy patient who suffers from cancer or before be exposed to carcinogen in heredity.The prevention effective dose can also refer to provide the amount of the preventive of preventative benefit in disease prevention.In addition, the prevention effective dose refers to separately or unites other medicament provides the preventive of preventative benefit in disease prevention amount for preventive of the present invention.

When being used for this paper, term " treatment " refers to eradicate, reduce or palliate a disease or the symptom of disease.In some embodiment, treatment refers to eradicate, remove, revise or control constitutional, regionality or metastatic carcinoma tissue by one or more therapeutic agents of administration.In certain embodiments, these terms refer to by one or more therapeutic agents of experimenter's administration of suffering from this disease are postponed the propagation of cancer or it reduced to minimum.

When being used for this paper, term " control (manage) " refers to by preventive that can not cure diseases to experimenter's administration or the beneficial effect that therapeutic agent obtains.In certain embodiments, one or more preventive of experimenter's administration or therapeutic agent are come " control " disease, thus prophylactic development or deterioration.

When being used for this paper, term " prevention " refers to the outbreak or the recurrence of one or more symptoms of prevention disease in the experimenter by administration preventive or therapeutic agent.

When being used for this paper, the undesired and disadvantageous effect that is produced by preventive or therapeutic agent contained in phrase " side effect ".Disadvantageous effect is always undesired, but undesired effect is not necessarily disadvantageous.The detrimental effect of preventive or therapeutic agent may be deleterious uncomfortable or dangerous.Chemotherapeutic side effect include but not limited to gastrointestinal toxicity such as early stage and form diarrhoea and flatulence, nauseating, vomiting, anorexia, leukopenia, anemia, neutrophil cell minimizing, weakness, abdominal colic, fever, pain late period, lose weight, dewater, bald head, dyspnea, insomnia, giddy, mucositis, xerostomia and renal failure, and constipation, N﹠M effect, temporary or permanent damage, influenza-like symptom, fluid retention and temporary or permanent sterile to kidney and bladder.Radiotherapeutic side effect includes but not limited to fatigue, xerostomia and forfeiture appetite.Biology, the side effect of therapy/immunotherapy included but not limited to the erythra of medicine-feeding part or swelling, influenza-like symptom (such as fever, shiver with cold and tired), digestive tract problem and anaphylaxis.The side effect of hormonotherapy includes but not limited to feel sick, fertility Issue, depression, forfeiture appetite, visual problem, headache and weight fluctuations.This area knows that also having a lot of other ill effects is that the patient often suffers, and for example sees " the desktop handbook of doctor physician " (thePhysicians ' Desk Reference), the 56th edition, and 2002, its complete being collected herein by reference.

4. accompanying drawing is described

Fig. 1 illustrates the IL-12 route of administration suppresses tumor growth to IL-12 influence.

Fig. 2 illustrates the IL-12 route of administration to carrying the influence of mice with tumor mortality rate.

Fig. 3 illustrates the influence (by facs analysis measure) of IL-12 route of administration to detected NK cell percentage among the DLN.

Fig. 4 illustrates ID and IM delivers the level of back RSV specific antibody in lung.This area graph shown the 3rd and 24 hours, the 1st, 2, the 3 and 4 weeks lung tissue of gathering by animal in the amount of detected antibody.

Fig. 5 illustrates that best IN delivers determining of volume and lung acquisition time in the Balb/c model.

Fig. 6 illustrates by splitting two kinds of possible bioavailability results that (splitting) dosage is realized.

Fig. 7 illustrates the synagis in the lung tissue lysate.

Fig. 8 has shown the result of synagis plaque check (synagis plaque assay).

Fig. 9 illustrates the decomposition diagram according to the needle assembly (assembly) of the present invention's design.

Figure 10 illustrates the partial cross section figure of Fig. 9 embodiment.

Figure 11 illustrates the injection device that Fig. 9 embodiment is installed on the syringe body and forms.

5. detailed Description Of The Invention

The invention provides the method that is used for one or more biologic activity agent (preferred therapeutic agents) of percutaneous drug delivery of experimenter, wherein the biologic activity agent is delivered to the intradermal zone of experimenter's skin.The present invention's part is found based on inventor's accident, promptly delivering pattern (comprising subcutaneous delivery and intramuscular delivery) with routine compares, when these therapeutic agents being delivered to the intradermal zone, they are delivered to the system of regional nodes rapidly, the benefit that provides this paper to announce thus.Although do not want to be subject to concrete mechanism of action, carry by the system of regional nodes in vivo according to the therapeutic agent that the inventive method is delivered, and enter circulation of systemic blood body and deep tissue environment.

The invention provides and be used to deliver improving one's methods of biologic activity agent, it especially provides following benefit, promptly absorb rapidly and enter regional nodes, institute's delivery of therapeutics agents improves i.e. deposition raising to the targeting of particular organization's (promptly carrying out a group or a confluent monolayer cells of specific function together), systemic bioavailability improves, tissue biological's availability improves, the deposition of medicament for the treatment of the predetermined of administration improves, organizing specific kinetics is improved, biological and pharmacodynamics (PD) rapid, and biological and pharmacokinetics (PK) is rapidly.These benefits of the inventive method are particularly useful for delivery of therapeutics agents.Intradermal delivery of therapeutics agents according to the inventive method is deposited on therapeutic agent in intradermal and the lymph zone, thereby causes therapeutic agent rapidly and biologically focusing on significantly in these zones.

The therapeutic agent of intradermal delivery has improved tissue biological availability in particular organization, include but not limited to mucous layer, skin histology, lymphoid tissue (for example lymph node), mucosal tissue, germinal tissue, cervical tissue, vagina tissue and form and the health any part of carrying out specific function is an organ, include but not limited to lung, spleen, colon, thymus by dissimilar tissues.In some embodiment, tissue comprises and environmental interaction or accessibility any tissue, for example skin, mucosal tissue.Other tissue that the present invention is contained includes but not limited to hemolymph sample system; Lymphoid tissue (for example relevant lymphoid tissue of epithelium and mucosa-associated lymphoid tissue or MALT (MALT can further be divided into the lymphoid tissue (D-MALT) of organized mucosa-associated lymphoid tissue (O-MALT) and disperse); Elementary lymphoid tissue (for example thymus and bone marrow); Secondary lymphoid tissue (for example lymph node, spleen, digestion, breathing and urogenital).Those skilled in the art will understand, and secondary MALT comprises gut associated lymphoid tissue (GALT); BALT (BALT); And urogenital system.MALT specifically comprises the peyer's patches in lymph node, spleen, tissue such as the jaw relevant with epithelial surface and nasopharynx tonsil, the small intestinal and the conjunctiva of the various joint knots in breathing and the genitourinary system, skin and eye.O-MALT comprises pharynx week lymph sample ring, the esophagus joint knot of tonsil (jaw, tongue, nose because of and pipe) and runs through the similar lymphoid tissue that whole digestive tract is scattered to anal canal by duodenum.

According to the inventive method delivery of therapeutics agents with identical reagent is delivered to subcutaneous (SC) or intramuscular zone and compares and cause the tissue biological availability to improve.Delivering reagent according to the inventive method, to make that tissue biological's availability improves when the delivery of therapeutics agents particularly useful, because it produces useful therapeutic effect, comprises and save dosage, improve effect of drugs and reduce side effect.

The therapeutic agent of delivering according to the inventive method is deposited in the intradermal zone, and at first is distributed in regional nodes's tissue of medicine-feeding part with high bioavailability, enters systemic circulation by propagating wider lymph delivery subsequently.In some embodiment, method of the present invention is effective especially for disease, disease or the infection of treatment deep tissue.

In some embodiment, the concentration that ID delivers back sedimentary biologic activity agent in particular organization is per about 5 nanograms of 50 microgram particular organizations (nanogram) medicaments; Per 50 microgram particular organizations, 10 piks (picogram) medicament; Per 50 microgram particular organizations 29 nanogram medicaments; Per 50 microgram particular organizations 10 pik medicaments are to the about 29 nanogram medicaments of per 50 microgram particular organizations; Per 50 microgram particular organizations 10 pik medicaments are to the about 150 nanogram medicaments of per 50 microgram particular organizations, or per 50 microgram particular organizations 10 pik medicaments.

The method that is used for intradermal delivery biologic activity agent (particularly therapeutic agent) is contained in the present invention, make with by the approach beyond the intradermal delivery (such as SC deliver, intramuscular is delivered, intravenous is delivered and the epidermis delivery) the delivery medicament compares, medicament has higher tissue biological's availability in particular organization.In some embodiment, the biologic activity agent (particularly therapeutic agent) of delivering according to the inventive method has and the similar bioavailability (comprising tissue biological's availability) of intravenous delivery medicament.

The method that is used for intradermal delivery biologic activity agent (particularly therapeutic agent) is contained in the present invention, makes to compare with medicament being delivered to deep tissue zone (for example SC), and medicament has higher tissue biological's availability in particular organization.Preferably, the biologic activity agent of delivering according to the inventive method is to show effect or develop or control disease and the therapeutic agent of administration includes but not limited to cancer (for example lymphoma, leukemia, breast carcinoma, melanoma, pulmonary carcinoma, renal carcinoma and colorectal carcinoma), transfer, tumor growth or infectious disease in order to treat, to prevent, postponing.

The method that is used for intradermal delivery biologic activity agent (particularly therapeutic agent) is contained in the present invention, makes to compare with medicament being delivered to deep tissue zone (for example SC), and reagent has higher therapeutic efficiency.In some embodiment, the biologic activity medicament of delivering according to the inventive method (particularly therapeutic agent) has the therapeutic efficiency similar to the intravenous administration medicament.

The present invention is contained and is used for the treatment of, the method for prevention or control disease, comprise with at least a therapeutic agent of predetermined close administration, wherein with by the conventional drug dose of delivering of other delivery approach (such as SC, IM and IV) compare, this predetermined close reduced at least half, at least 5 times, at least 10 times.

In some embodiment, the present invention is encompassed in the method for treatment among the needed human experimenter, prevention or control cancer, cancerometastasis or tumor growth, comprises the ID zone that therapeutic agent is delivered to human experimenter's skin.The intradermal delivery medicament causes comparing the bigger reduction of tumor growth with the identical medicament of delivering by ID in addition of approach (for example SC, IM, IV, epidermis) delivery in order to treat, prevent or control cancer, cancerometastasis or tumor growth.In some embodiment, the intradermal delivery medicament causes delivering the life-span intermediate value prolongation that this medicament is compared the human experimenter with the approach in addition of delivering by ID in order to treat, prevent or control cancer, cancerometastasis or tumor growth.

Direct targeting intradermal zone according to instruction of the present invention makes that the effect of these medicaments begins rapider and bioavailability (comprising tissue biological's availability) is higher for the routine of these biologic activity agent (comprising therapeutic agent) is delivered pattern (comprising subcutaneous delivery).The inventor finds, the medicament of delivering according to the inventive method can arrive the controlled intradermal administration of corium blood vessel and lymphatic capillaries and absorb also whole body distribution rapidly by selectivity, make and compare that this medicament can be brought into play their beneficial effect more rapidly with conventional mode of administration (such as subcutaneous administration).

When being used for this paper, be delivered in the intradermal zone or intradermal delivery means the biologic activity agent is administered into mode in the corium, the mamillary corium that makes medicament be easy to arrive to be rich in vascular, and absorb rapidly in blood capillary and/or the lymphatic vessel and to become the whole body biology available.This can be achieved like this, and being about to medicament, to place the upper area of corium be mamillary corium, and perhaps the top of the less reticular corium of vascular makes reagent be easy to be diffused in the mamillary corium.The biologic activity agent below mamillary corium and in reticular corium but enough be higher than controlled delivery in this dermal zone at interface between corium and the subcutaneous tissue should make medicament effectively (outwards) move to (undisturbed) vascular and lymphatic capillaries bed (in mamillary corium), it can absorb by these capillary tubies and enter the body circulation at this, and can not detained by any other skin histology zone in current process.In some embodiment, with the biologic activity agent mainly place at least about 0.3mm, more preferably at least about 0.4mm and most preferably at least about the degree of depth of 0.5mm to being no more than about 2.5mm, will causing the rapid absorption of medicament more preferably no more than the about 2.0mm and the degree of depth that is most preferably not exceeding about 1.7mm.Although do not want to be subject to particular mechanism of action, the bottom that the biologic activity agent is mainly placed the bigger degree of depth and/or enter reticular corium may cause lymph effective inadequately to the absorption of medicament, because medicament will slowly absorb in less reticular corium of vascular or subcutaneous area.

5.1 the method for intradermal administration

The present invention is contained and is used for therapeutic agent described herein and illustrative or the material intradermal delivery method to the intradermal zone of experimenter's skin, preferred selectivity and selectively targeted intradermal zone but not pass it.In a most preferred embodiment, direct targeting intradermal zone.Remain the preparation of delivery of therapeutics agents in case prepared to contain, usually preparation is transferred to and be used for the injection device that deliver in the intradermal zone, for example syringe.By specificity and selectivity (directly preferred) targeting intradermal zone, deliver improved treatment and the clinical effectiveness that preparation of the present invention provides the medicament that is better than conventional delivery pattern (comprising IM and SC) according to method of the present invention.Intradermal delivery method of the present invention provides benefit and improvement, includes but not limited to the pharmacokinetics improved, absorbs rapidly to enter regional nodes, and the targeting of particular organization is improved and tissue biological's availability is improved.Method of the present invention causes pharmacokinetics to be improved, and is improved such as the absorption that enters the intradermal zone.Preparation of the present invention can be delivered to intradermal space with the form of injecting (bolus) or infusion.

The method that is used for intradermal delivery biologic activity agent (particularly therapeutic agent) is contained in the present invention, make with by the approach beyond the intradermal delivery (such as SC deliver, intramuscular is delivered, intravenous delivery and epidermis delivery) deliver this medicament and compare, this medicament has higher tissue biological's availability in particular organization.In some embodiment, the biologic activity agent (particularly therapeutic agent) of delivering according to the inventive method has to intravenous delivers the similar bioavailability of this medicament (comprising tissue biological's availability).

The method that is used for intradermal delivery biologic activity agent (particularly therapeutic agent) is contained in the present invention, makes to compare with this medicament being delivered to darker tissue regions (for example SC), and this medicament has higher tissue biological's availability in particular organization.Preferably, the biologic activity agent of delivering according to the inventive method is for treatment of diseases, prevention, delay outbreak or progress or control disease and the therapeutic agent of administration, and described disease includes but not limited to cancer (for example lymphoma, leukemia, breast carcinoma, melanoma, pulmonary carcinoma, renal carcinoma and colorectal carcinoma), transfer, tumor growth or infectious disease.

The method that is used for intradermal delivery biologic activity agent (particularly therapeutic agent) is contained in the present invention, makes to compare with this medicament being delivered to darker tissue regions (for example SC), and this medicament has higher therapeutic efficiency.In some embodiment, the biologic activity agent (particularly therapeutic agent) of delivering according to the inventive method has to intravenous delivers the similar therapeutic efficiency of this medicament.

The present invention is contained and is used for the treatment of, the method for prevention or control disease, comprise with at least a therapeutic agent of predetermined close administration, wherein compare with deliver the conventional drug dose of delivering of approach (such as SC, IM and IV) by other, this predetermined close reduced at least half, at least 5 times, at least 10 times.

In some embodiment, the present invention is encompassed in the method for treatment among the needed human experimenter, prevention or control cancer, cancerometastasis or tumor growth, comprises the ID zone that therapeutic agent is delivered to human experimenter's skin.The intradermal delivery medicament causes comparing the bigger reduction of tumor growth with the identical medicament of delivering by ID in addition of approach (for example SC, IM, IV, epidermis) delivery in order to treat, prevent or control cancer, cancerometastasis or tumor growth.In some embodiment, the intradermal delivery medicament causes delivering the life-span intermediate value prolongation that medicament is compared the human experimenter with the approach in addition of delivering by ID in order to treat, prevent or control cancer, cancerometastasis or tumor growth.

According to the present invention, directly the intradermal administration for example can be used based on any other means in the injection of microneedle and infusion system or the accurate targeting intradermal zone that those skilled in the art will know that and finish.Concrete device comprises the U.S. Patent number 6 that disclosed WO02/02179 of disclosed WO01/02178 on January 10th, 2002, on January 10th, 2002, December in 2002 were issued on the 17th, 494, the U.S. Patent number 6 that on May 27th, 865 and 2003 issued, 569, disclosed device in 143 (all these complete being collected herein by reference), and illustrative device among Fig. 9-11.Methodology and device based on miniature sleeve pipe and microneedle also are described in the U. S. application series number of submitting on June 29th, 2,000 09/606,909 (its complete being collected herein by reference).The steel sleeve pipe of standard also can use the apparatus and method of describing in the U.S. serial of submitting on October 14th, 1,999 417,671 (complete being collected herein by reference) and be used for intradermal delivery.These method and apparatus are delivered medicament by the narrow specification (30G or narrower) " miniature sleeve pipe " with limited penetration depth (scope is 10 μ m-2mm normally), and this is limited by telescopic total length or hub (hub) parts (feature) the sleeve pipe total length in addition that is exposed to controlling depth.

The experimenter of intradermal delivery of the present invention is a mammal, preferred people.The biologic activity agent of delivering according to the inventive method can be delivered in the intradermal zone by pin or the sleeve pipe of the about 5mm of the normally about 300 μ m-of length.Preferably, pin or telescopic length are the about 1mm of about 300 μ m-, and the degree of depth of outlet insertion experimenter skin is 1mm-3mm.Preferably, use the pin or the sleeve pipe of small dimension, promptly 30-36 number, preferred 31-34 number.The degree of depth of 0.3mm (300 μ m)-1.5mm is preferably inserted in pin or telescopic outlet.

According to the present invention, directly the intradermal administration for example can be used realizing based on the injection of microneedle and infusion system or any other means that those skilled in the art will know that of accurate targeting intradermal zone.Concrete device comprises the U.S. Patent number 6 that disclosed WO02/02179 of disclosed WO01/02178 on January 10th, 2002, on January 10th, 2002, December in 2002 were issued on the 17th, 494, the U.S. Patent number 6 that on May 27th, 865 and 2003 issued, 569, disclosed device in 143 (all complete being collected herein by reference), and illustrative device among Fig. 9-11.Methodology and device based on miniature sleeve pipe and microneedle also are described in the U. S. application series number of submitting on June 29th, 2,000 09/606,909 (complete being collected herein by reference).The steel sleeve pipe of standard also can use the apparatus and method of describing in the U.S. serial of submitting on October 14th, 1,999 417,671 (complete being collected herein by reference) and be used for intradermal delivery.These method and apparatus are delivered medicament by the narrow specification (30G or narrower) " miniature sleeve pipe " with limited penetration depth (scope is 10 μ m-2mm normally), and this is limited by telescopic total length or the sleeve pipe total length that is exposed to beyond the boss assembly of controlling depth.

The method of intradermal administration comprises any other means based on the injection of microneedle and infusion system or accurate targeting intradermal space.The method of intradermal administration not only contains the injection means based on micro device, also contain other delivering method, such as make that fluid or powder enter intradermal space few with pin or without the graphic intradermal injection of impact injection, the awns of pin, by the enhanced iontophoresis of micro device and fluid, solid or other dosage form the direct deposition in skin.

In a specific embodiment, by the graphic injection of intradermal awns preparation of the present invention is administered in the intradermal zone of experimenter's skin, see for example Flynn etc., 1994, Chest106:1463-5 (complete being collected herein by reference).In a specific embodiment, illustrative method is delivered to preparation of the present invention in the intradermal zone of experimenter's skin below using.With the preparation inhalation syringe, the 1ml that for example is furnished with No. 20 pins does not have latex (latex free) syringe; After device to be injected is filled, change No. 30 pins that are used for the intradermal administration into.Make progress near experimenter's (for example mice) the skin and the inclined-plane of pin with the most shallow as far as possible angle, and skin is collapsed tightly.Slowly promote volume injected in the time of second and form typical " blister (bleb) " at 5-10 then, slowly extract pin subsequently.Preferably, only use an injection site.More preferably, volume injected is no more than 100 μ l, and this part is because big volume injected may increase the fact of overflowing and entering organization space (for example subcutaneous space) on every side.

Conventional entry needle, sleeve pipe or the microneedle that all known types are contained in the present invention separately or the purposes of using in the spininess array.Term " pin " is intended to contain all these pin spline structures when being used for this paper.When these structures were cylinder in essence, term " microneedle " was intended to contain less than about No. 30 structure normally about 31-50 number when being used for this paper.Therefore, the non-cylinder structure that term " microneedle " is contained will have equivalent diameter, and comprise pyramid, rectangle, octagonal, wedge shape and other geometry.

Impact type (ballistic) fluid injector, powderject formula delivery apparatus, piezoelectricity, electronic, the auxiliary delivery apparatus of electromagnetism, the auxiliary delivery apparatus of gas are contained in the present invention, their direct transdermals and preparation of the present invention directly is delivered to target position in the corium space.

With the practical methods of preparation targeting intradermal space of the present invention and non-key, as long as the skin that it penetrates the experimenter arrives the desired target degree of depth of intradermal space but not passes it.The actual optimal penetration degree of depth will change along with the thickness of experimenter's skin.In most applications, the degree of depth of transdermal is about 0.5-2mm.No matter concrete intradermal delivery apparatus and method, method of the present invention preferably with preparation targeting of the present invention at least the degree of depth of 0.5mm more preferably no more than 2.0mm, be most preferably not exceeding 1.7mm to the degree of depth that is no more than 2.5mm.In some embodiment, be delivered to just preparation below horny layer and comprise the target degree of depth of epidermis and upper part of dermis, for example about 0.025mm is about 2.5mm extremely.For the specific cells in the targeting skin, the preferred target degree of depth depends on the concrete cell of targeting and the thickness of concrete experimenter's skin.For example, for youth's lattice Han Shi (Langerhan) cell in the targeting human skin corium space, deliver near small part and need comprise epidermal tissue's degree of depth, the normally about 0.025mm of the scope in human body is to about 0.2mm.

Preparation according to delivery of the present invention or administration comprises that their solution, suspension, gel, granule (such as miniature and nano-particle, or suspending or dispersion) and original positions thereof in pharmacopedics acceptable diluent or solvent form carrier.

In other embodiment preferred, present invention resides on experimenter's the skin and select the injection site, and the injection site on the cleaning experimenter skin, and then discharge biologic activity agent (particularly therapeutic agent) by delivery apparatus and enter experimenter's skin.In addition, this method comprises with biologic activity agent of the present invention (particularly therapeutic agent) and is full of delivery apparatus.In addition, this method comprises compressing skin, make limiter (limiter) part (engaging) experimenter's the skin of fitting, and exert pressure, thereby collapse tight experimenter's skin, and behind injection medicament by extracting needle cannula in the skin out.Also have, the step that front tip is inserted skin also is defined as the degree of depth that front tip inserts skin by about 1.0mm about 2.0mm extremely, most preferably inserts the skin depth of 1.5mm+0.2 to 0.3mm.

In a preferred embodiment, with front tip insert zoodermic step also be defined as angle that front tip inserts skin usually at about 15 degree with interior perpendicular to skin, most preferably become 90 degree with skin with interior, and also be defined as normally vertical with respect to the fixed angle orientation of skin attachement face at about 5 degree.In a preferred embodiment, limiter centers on needle cannula, and has the flat surfaces of applying skin surface usually.Also have, delivery apparatus comprises having tube and be contained in piston in the tube, and piston can depress, thereby is discharged the syringe of medicament by delivery apparatus by the front tip of needle cannula.

In a preferred embodiment, discharge biologic activity agent (particularly therapeutic agent) by delivery apparatus and also be defined as the forefinger pressure piston that holds hypodermic needle and use the another hands with, and by holding hypodermic needle and press the piston on the hypodermic needle to discharge medicament with one, and front tip is inserted zoodermic step also be defined as the skin of oppressing animal with limiter by delivery apparatus with the thumb of another hands.In addition, this method also comprises loads onto the step of needle assembly (assembly) with the tube point of syringe, and wherein needle assembly comprises needle cannula and limiter, and can be included in to the tube point and load onto a step that exposes the tube point before the needle assembly by the medicated cap of removing it.Perhaps, the step that the front tip of pin is inserted experimenter's skin can be defined as with one and hold hypodermic needle, thereby the skin with limiter compressing animal collapses the skin of tight animal simultaneously, and discharges medicament or discharge medicament by the depress piston with this hands by depress piston with the forefinger of this hands.This method also is included in is extracted drug injection back in experimenter's skin the front tip of needle cannula out by experimenter's skin.Also have, this method comprises inserts the skin depth of preferred about 1.0mm to about 2.0mm, the most preferably degree of depth of 1.5mm+0.2 to 0.3mm with front tip.

Some feature of finding the intradermal medication provides useful clinically PK/PD and dose accuracy.For example, find the pin outlet is placed appreciable impact PK/PD parameter in the skin.Have the routine on inclined-plane or the outlet of standard specifications pin and have bigger exposure height (vertical height of outlet).Although needle point can be placed the desired depth in intradermal zone, the medicament that the exposure height of pin outlet causes more greatly delivering is deposited on from the nearer much shallow degree of depth of skin surface.As a result, because the counter-pressure that applies of skin self and by the pressure of the flow volume accumulation of injection or infusion, medicament is tending towards being flowed out by skin, and infiltrates the lower skin area of pressure, such as subcutaneous tissue.That is to say that the pin outlet that has than big exposure height will effectively seal in the big degree of depth, and the outlet with identical exposure height can not seal effectively when placing the more shallow degree of depth in intradermal zone.Usually, the exposure height of pin outlet is 0 to about 1mm.The pin outlet of exposure height 0mm does not have the inclined-plane, and is positioned at needle point.In this case, the degree of depth of outlet is identical with the penetration depth of pin.Be all to have measurable exposure height by the inclined-plane or by the pin outlet that the pin lateral opening forms.Be appreciated that one piece of pin can have more than one opening or the outlet that is suitable for medicament is delivered to dermal zone.

Find that also the pressure by control injection or infusion can overcome the high counter-pressure that applies in the ID administration process.By directly constant pressure being placed on the liquid surface, can realize more constant delivery speed, this may make and absorb optimization and obtain improved pharmacokinetics.Can also control and deliver speed and volume, thereby position formation pomphus (wheal) is being delivered in prevention, and the device that prevents counter-pressure will arrive corium is released skin and/or entered subcutaneous area.Only use ordinary skill just can determine to obtain the suitable delivery speed and the volume of these effects by experiment.The distance that increases between many pieces of pins is allowed wider liquid distribution and is improved and deliver speed or increase fluid volume.

Can be used for carrying out medication of the present invention comprises by injecting (bolus) and the infusion dual mode is delivered the biologic activity agent to human or animal experimenter.Bolus dose is the single dosage of delivering in single volume unit (being no more than about 10 minutes usually) in the short period.The infusion administration is included in the long period (usually above about 10 minutes) with set rate (can be constant or variable) administration fluid.In order to deliver medicament, the device that arrives corium is placed near experimenter's skin, provide, and medicament is delivered or is administered in the intradermal zone by the path in the intradermal zone of direct targeting, they can play a role the part at this, are perhaps absorbed and the whole body distribution by blood flow.The device that arrives corium can connect the container of waiting to deliver medicament is housed.

The generation that is entered the delivery in intradermal zone by container can be passive, promptly need not to treat the delivery medicament and applies external pressure or other driving means, and/or be initiatively, promptly exerts pressure or other driving means.The example that produces the preferred means of pressure comprises that pump, syringe, pen (pen), elastomer film, air pressure, piezoelectricity, electronic, electromagnetism or infiltration aspirate or Bei Laiweier (Belleville) spring or packing ring or its combination.If desired, the delivery speed of medicament can be the variable control that is subjected to producing the means of pressure.

In some embodiment, the method for pharmacokinetics of controlling the biologic activity agent of institute's administration by the advantage of uniting two or more zones that are delivered in the skin or the degree of depth is contained in the present invention.Particularly, the invention provides biologic activity agent described herein (particularly therapeutic agent) is delivered to shallow SC and ID zone to realize mixing the method for PK characteristic, promptly the PK that delivers of a part and ID is similar and another part is similar to the PK of SC delivery.This provides fast and high peak value outbreak level and lower prolongation cyclical level for activating agent biology (particularly therapeutic agent).These methods are disclosed in the U. S. application series number of submitting on May 6th, 2,003 10/429,973 (complete being collected herein by reference).In some embodiment, biologic activity agent (particularly therapeutic agent) is delivered to the one or more positions that comprise two or more zones.In other embodiments, biologic activity agent (particularly therapeutic agent) is delivered to a plurality of positions, each comprises one or more zones.

Method of the present invention contains the controlled delivery of the biologic activity agent (particularly therapeutic agent) of using following algorithm, and the logic composition that described algorithm had comprises the pharmacokinetics model of physiological mode, rule-based model or moving average method, therapy, algorithm, predictive control model or its combination of monitor signal processing.

Method of the present invention contains the shallow SC of associating and ID delivers the method that realizes improved PK result.Only use wherein a kind of delivery service area or another kind can not realize these results.Multi-section position deposition via correct device construction and/or medication can obtain uniqueness and useful result.Basic know-why is that the PK result that microneedle is delivered is special for the fluidic deposition degree of depth of institute's administration and pattern, and making can be by device design and engineering or by such as making the ID zone cross technology such as carrying object and this deposition of machinery control.

In addition, the present invention includes length less than 5mm, be used for hypodermic pin (miniature or other form).The shallow SC that arrives about 3mm degree of depth delivers and produces and the dark SC PK much at one that uses conventional art.Never the effect that adopts shallow SC to deliver separately produces more in check characteristic.In fact, think that the previous degree of depth less than 5mm is not in the SC zone.

Mix delivery (no matter being) and cause two-phase or hybrid dynamics feature by device design or technology.The faint difference of device length (1mm to 2mm to 3mm) 30 produces significant difference in PK result.Use and usually to suppose that the pin length that the pin end is arranged in the ID zone can obtain SC sample characteristic.The PK result that shallow SC delivers delivers more the unification of making peace than standard SC.The restriction of the target tissue degree of depth especially is subjected to the control of exposure height (vertical height), administration volume and the medicine-feeding rate of the degree of depth that the outlet of pin or sleeve pipe inserts, outlet.Those skilled in the art need not undo experimentation just can determine suitable parameters.

5.1.1 be used for the device of intradermal administration

The U.S. Patent number 6 that uses the WO02/02179 that announces in that this area is known or the WO01/02178 that on January 10th, 2002 announced, on January 10th, 2002, December in 2002 to issue in 17th, 494, the U.S. Patent number 6 that on May 27th, 865 and 2003 issued, disclosed any apparatus and method are come administration biologic activity agent of the present invention (comprising therapeutic agent) in 569,143 (all complete being collected herein by reference).

Preferably, be used for having the structural means that the control transdermal arrives the intradermal space desired depth according to the device that method of the present invention is carried out the intradermal administration.This is modal to be to realize that by the widened section or the hub that link to each other with the axle (shaft) that arrives the corium device it can be taked gasket construction (backing structure) or be used to install the form of the platform of pin.Length as the microneedle that arrives the corium device is easy to change in manufacturing process, and is made into length usually less than 2mm.Microneedle is still very sharp-pointed, and specification is very little, thereby further alleviates pain and other sensation in injection or the infusion process.They can be used as one single chamber microneedle and are used for the present invention, perhaps can or be made into linear array or two-dimensional array with the assembling of many pieces of microneedle, thus improve delivery speed or the amount of the material of delivering at the appointed time.Can be by end, side or the two its material of ejaculation of pin.Microneedle can be added multiple device, such as holder that also can be used for limiting penetration depth (holder) and shell (housing).The device of arrival corium of the present invention can also add the container that material is housed before delivery, perhaps pump or be used under pressure delivering other device of medicine or other material.Perhaps, hold the device that corium arrives device and can outsidely connect these additional assemblies.

The intradermal medication comprises based on the injection of microneedle and infusion system or accurate spatial any other means of targeting Intradermal.The intradermal medication not only contains the injection means based on microneedle, also contain other delivering method, such as making fluid or powder enter the few of intradermal space with pin or without the impact injection of pin, by the enhanced iontophoresis of micro device, and fluid, solid or other dosage form direct deposition in skin.

In some embodiment, the invention provides the delivery apparatus that comprises needle assembly, is used to carry out the intradermal injection.Needle assembly has the adapter (adapter) that enables to be installed on the container (such as syringe etc.) that can be pre-charged with.Needle assembly is by the adapter support, and has the hollow body that front end is stretched out by adapter.Limiter centers on pin, and stretches out the front end that points to pin by adapter.Limiter has the skin attachement face of the skin of the animal (such as the people) that is suitable for fitting.The front end of pin stretches out selected distance by the skin attachement face, makes limiter restriction pin can penetrate the zoodermic amount or the degree of depth.

In a specific embodiment, the hypodermic needle assembly that is used for the inventive method comprises that being used for execution relates to the element essential to the invention of improving one's methods that biologic activity agent (comprising therapeutic agent) is delivered to experimenter's skin (preferred human experimenter's skin), may further comprise the steps: the delivery apparatus that comprises needle cannula is provided, described needle cannula comprises forward tip, and the contained medicament of needle cannula and delivery apparatus carries out fluid communication, described device also comprises the limiter part around needle cannula, and limiter partly comprises the skin attachement face, the needle point of needle cannula is partly stretched out by limiter, the distance that surpasses the skin attachement face equals about 0.5mm to about 3.0mm, and needle cannula has the planar fixed angle orientation with respect to limiter part skin attachement face; Needle point is inserted the skin of animal and with limiter skin attachement face applying skin surface partly, makes the skin attachement face restriction needle guard tip of limiter part penetrate into zoodermic skin corium; And enter animal skin by drug delivery device ejected matter by the needle guard tip.

In a specific embodiment, disclosed drug delivery device among Fig. 9-10 is contained in the present invention, their illustrations can be used for putting into practice the example of the drug delivery device of the inventive method that is used for intradermal injection.Illustrative device 10 comprises the needle assembly 20 that can be installed on the syringe cylinder 60 among Fig. 9-10.The delivery apparatus of operable other form comprises U.S. Patent number 5,279,586, disclosed various pen among U.S. Patent Application Serial 09/027,607 and the PCT application number WO00/09135 (its content intact is collected herein by reference).

Needle assembly 20 comprises the hub 22 of supporting needle cannula 24.Limiter 26 holds at least a portion hub 22, make limiter 26 usually around needle cannula 24, sees as clear among Fig. 9.

The accepter 32 of the enough syringes of one end, 30 energy of hub 22 is fixing.Be used to hold according to the multiple injector type of the material of intradermal delivery of the present invention and can use, hereinafter provided several examples with the needle assembly of design.The other end of hub 22 preferably includes the extension 34 of interface (abutment surface) 36 nested acceptance in the limiter 26.Preferably on limiter 26, provide a plurality of ribs (rib) 38 structural intergrity to be provided and to be convenient to operate needle assembly 20.

By the size of suitable design component, can firmly control the front end of pin 24 or the distance ' ' d ' ' between the skin attachement face 42 on point 40 and the limiter 26.Distance ' ' d ' ' preferably at about 0.5mm to the scope of about 3.0mm, 1.5mm ± 0.2mm to 0.3mm most preferably from about.The distance that extends beyond skin attachement face 42 when the front end 40 of needle cannula 24 is in this scope the time, and the intradermal injection is protected, because pin is can not the typical animal skin corium of penetration ratio darker.Usually, outer skin is that the thickness of epidermis is the 50-200 micron, and corium promptly the thickness of inner thicker skin layer be 1.5-3.5mm.Be subcutaneous tissue (being also referred to as hypodermal layer sometimes) and muscular tissue successively below the skin corium.

Can see that limiter 26 comprises opening 44 by Fig. 9 is clear, the front end 40 of needle cannula 24 is by wherein protruding.Can needs as the case may be control the spatial relationship between opening 44 and the front end 40.In exemplary embodiment, skin attachement face 42 normally planar or smooth and successive, thereby can be at the stable needle assembly 20 of settling of animal skin.Although there is not particular instantiation, make normally smooth skin attachement face 42 comprise or the bossing of rib form or the female of groove form with enhanced stability or to be convenient to the pin covers to needle point 40 are installed may be favourable.In addition, can extend beyond the plane of skin attachement face 42 along limiter 26 lateral ribs 38.

No matter the shape and the profile of skin attachement face 42, embodiment preferred comprise enough normally planar or smooth, with contact skin so that make the surf zone that syringe is stable with respect to experimenter's skin.In most preferred arrangement, skin attachement face 42 is convenient to syringe is maintained with respect to the normally vertical orientation of skin surface, and is convenient to exert pressure at skin in injection process.Thus, in preferred embodiments, the size of limiter or external diameter are 5mm at least.Main size will depend on to be used and the packing restriction, but diameter is less than 15mm or bigger easily, preferred 11-12mm.

Must note, although Fig. 9 and 10 illustrations contain the assembly (two-pieceassembly) of two parts, wherein hub 22 is what to separate with limiter 26, is not limited to this arrangement yet be used for device of the present invention.Form hub 22 and limiter 26 is alternatives of Fig. 9 and 10 example illustrated by single piece of plastic material is whole.In addition, might hub 22 be fixed on the illustrated position of Figure 10 on the limiter 26, make needle assembly 20 after assembling, become single-piece part by adhesion or other method.

Hub 22 and limiter 26 have advantage aspect the reality manufacturing intradermal pin.Preferred pin size is small size hypodermic needle, usually is called No. 30 or No. 31 pins.This minor diameter pin is given and is done pin to such an extent that thereby the enough short skin corium that prevents overpenetration to surpass animal has brought challenge.Limiter 26 and hub 22 total lengths easy to use penetrate the much bigger pin 24 of effective pin length of individual tissue than in injection process.By needle assembly according to this paper design, make and obtained reinforcement because can make and assembling process in operate longer pin and still obtain hour hand in the advantage of finishing aspect the intradermal injection.

Figure 11 illustration needle assembly 20 be fixed on the medicament reservoir (such as syringe 60), thereby form device 10.Normally cylindrical syringe body 62 can be made by plastics or glass, as road known in the art.Syringe body 62 provides to be used for containing at injection process and has remained the container 64 of administration material.One end of piston rod 66 has manual activation flange 68, and the other end has braking thing 70, as road known in the art.Piston rod 66 passes the manually mobile of container 64 forces the material in the container 64 to be discharged by the end 40 of pin, as what expect.

Can hub 22 be fixed on the syringe body 62 with multiple known way.In an example, between the outside of the inside of hub 22 and syringe body 62 port of export parts 72, provide interference engagement (an interference fit).In another example, provide conventional Luer to cooperate (Luer fit) to arrange, thereby hub 22 has been fixed on the end of syringe 60.Can be understanded by Fig. 9-11, this needle assembly of design is easy to adapt to multiple conventional syringe type.

The invention provides the intradermal needle injection that is applicable to multiple injector type.Therefore, the present invention is being convenient to have significant advantage aspect large-scale manufacturing of the mode of economy and assembling intradermal pin.

Before inserting needle cannula 24, the injection site on selection and the clean animal skin.After selecting and cleaning the position, with the skin of 90 angles of spending, touch skin usually until skin attachement face 42 with the front end 40 insertion animals of needle cannula 24.Skin attachement face 42 stops needle cannula 42 to pass dermal layer of the skin and material is injected hypodermic layer.

After needle cannula 42 is inserted skin, material is injected intradermal.Material can be pre-filled in the syringe 60, or just fills before injection and store and wherein or only fill before injection.According to individuality preference and injector type, carry out the method for injection and can take several variations.In any situation, penetrating of needle cannula 42 is most preferably not exceeding about 1.5mm, because skin attachement face 42 stops and anyly further penetrates.

Also have, in the process of intradermal injection administration, the front end 40 of needle cannula 42 inserts the skin corium of skin, produces the counter-pressure of reasonable amount in the process of injected material.This counter-pressure can be in the rank of 76psi.In order to allow user that the power that syringe piston rod 66 applies necessary minimum is just reached this pressure, the syringe cylinder 60 of preferred little internal diameter is such as 0.183 " (4.65mm) or littler.Thus, method of the present invention comprises selects the syringe be used to inject, when injecting by the syringe ejected matter in its footpath width be enough to produce the strength that is enough to overcome the skin corium counter-pressure.

In addition, because the intradermal injection is carried out with small size material to be injected usually, promptly be no more than the rank of 0.5ml, preferred about 0.1ml, so the syringe cylinder 60 of preferred little internal diameter, thereby minimum is reduced in the dead band (dead space) that may cause injecting the waste material of catching between braking thing 70 and the syringe shoulder after finishing.Also have, because the volume of material is little, the rank of 0.1ml, thus the preferred little syringe cylinder of internal diameter, thus levels of substance in the process of braking thing will be inserted and the gas headspace of braking between the thing 70 is reduced to minimum.In addition, little internal diameter has strengthened the ability of checking and manifest material volume in the syringe cylinder.

5.2 according to the inventive method treatment disease

The present invention contain for prevent, treat improvement and disease, disease or infect one or more relevant symptoms by the ID zone that medicament is delivered to experimenter's skin to animal, preferred mammal, optimum is chosen and is used one or more therapeutic agents.Method of the present invention is for treatment or prevent lymphoid disease or disease, constitutional or metastatic neoplastic diseases (being cancer) and infectious disease particularly useful.That can know as this area or can accept compositions with pharmacopedics as described herein or preparation provides therapeutic agent.

The present invention is encompassed in the method for treatment among the needed experimenter, prevention or control disease or disease, and described method comprises that one or more therapeutic agents that will treat effective dose or prevention effective dose to described experimenter are administered to the intradermal zone of experimenter's skin.

The invention provides by therapeutic agent being delivered to experimenter's intradermal zone, make and compare that this therapeutic agent is more effective, and in the experimenter, treat or prophylactic method with conventional delivery approach (for example IM, IV or SC).

In some embodiment, the method that is used at experimenter's treatment or prophylaxis against infection diseases is also contained in the present invention, comprises one or more therapeutic agents that combine with infective agent or its cell receptor of drug treatment or prevention effective dose.Can cause by infective agent by the infectious disease of molecular therapy of the present invention or prevention, include but not limited to virus, antibacterial, fungus and protozoacide.

Especially provide following benefit according to intradermal delivery biologic activity of the present invention agent, promptly absorbed rapidly and enter regional nodes, institute's medicament of delivering is to the targeting and the deposition raising of particular organization, and system and tissue biological's availability raising.These benefits are particularly useful such as therapeutic agents such as antitumor agent, antibody and antibiotic for delivering.Intradermal delivery medicament according to the inventive method is deposited on medicament in intradermal and lymph zone and the deep tissue, causes medicament rapidly and in biologically focusing on these zones and tissue significantly.

With the direct targeting lymph of various therapeutic agent medicine entities, realized useful therapeutic effect by intradermal delivery, comprised and save dosage, improve effect of drugs, reduce side effect, reduce metastatic potential and prolong life cycle.The invention provides and be used for the treatment of improving one's methods of disease, delivering method promptly of the present invention makes therapeutic agent not only whole body but also local deposits.As a result, the medicament deposition by improving sensitivity, institute's delivery of therapeutics agents, tissue biological's availability, begin effect and removing faster faster, the invention provides and be used for the treatment of improving one's methods of disease (such as cancer).The invention provides in order to treat the method for disease (particularly cancer) at least a therapeutic agent of administration, comprise with controllable rate, volume and pressure medicament is delivered to the intradermal zone of experimenter's skin, make medicament be deposited in the ID zone and and absorb by the lymph vascular system.

Medicament deposition by improving sensitivity, institute's delivery of therapeutics agents, tissue biological's availability, begin effect and removing faster faster, the present invention also provides and has been used for the treatment of improving one's methods of the disease that is positioned health particular organization and organ, such as the infection of those tissues and organ, for example respiratory tract infection.The invention provides in order to treat the method for disease (particularly infecting) at least a therapeutic agent of administration, comprise with controllable rate, volume and pressure medicament is delivered to the intradermal zone of experimenter's skin, make medicament be deposited in the dermal zone and and absorb by the lymph vascular system.

In a specific embodiment, the invention provides and be used to deliver improving one's methods of antitumor agent, chemotherapeutant, antibody, antiangiogenic agent, antiinflammatory, immunotherapeutic agent etc., its clinical efficacy and therapeutic effect are improved.The routine treatment of primary cancer damage is finished by former piece of excision, local radiotherapy kill tumor tissue or whole body administration antitumor drug kill tumor usually.Preceding two kinds of methods have local more on the scope and can potentially will reduce to minimum advantage to the injury of non-target organ when can be used for treating.On the contrary, because this treatment is partial, so influence has broken away from primary tumo(u)r and has been positioned transitional cell limited in one's ability of other tissue.General antitumor agent therapy has the tumor cell that influences disperse and the ability of primary tumo(u)r, but this whole body is delivered the ability that has increased injury healthy organ, tissue and system.

Can directly arrive vein and two kinds of networks of lymph of corium according to intradermal delivery of therapeutics agents of the present invention (such as antibody and antitumor agent), and unique systemic drug kinetic effect is provided.By arriving near these networks of target (such as tumor), just may realize treating benefit.At first, will be enhanced with respect to the general toxicity local action of adverse events because the antitumor agent of intradermal delivery place physically tumor near, cause dosage to reduce and the side effect reduction.Equally because tumor mainly utilizes the whole body vascular system to shift conveying, so the antitumor agent of intradermal delivery can targeting these break away from cells, and reduce the ability that shifts.The antitumor agent of intradermal administration can also arrive immunocyte by the mediation of lymph network, influences the maturation of immunocyte and the conveying that anticancer cell (T, B, NK, macrophage etc.) arrives tumor locus.In addition, the antitumor agent of intradermal delivery will cause general to distribute, and provide the position of the extensive disperse of arrival and the added advantage of organ in the mode similar to many current therapy.The invention provides improving one's methods of the bioavailability of enhancing biologic activity agent in particular organization, include but not limited to lymph, mucosa, lung, spleen, thymus or colon.Especially provide following benefit according to intradermal delivery biologic activity of the present invention agent, promptly absorbed rapidly and enter regional nodes, institute's medicament of delivering is to the targeting and the deposition raising of particular organization, and whole body and tissue biological's availability raising.These benefits are particularly useful such as therapeutic agents such as antitumor agent, antibody and antibiotic for delivering.Intradermal delivery medicament according to the inventive method is deposited on medicament in intradermal and lymph zone and the deep tissue, causes medicament rapidly and in biologically focusing on these zones and tissue significantly.

The intradermal therapy may have bigger benefit to some cancer type.Peripheral cancer or highly be confined to, be present in or the cancer relevant with the purpose vascular system can be enjoyed the largest benefit of this class therapy.The concrete cancer of enjoying unusual benefit include but not limited to melanoma and other skin carcinoma (sarcoma etc.), lymphoma or adenoid other cancer, breast carcinoma or periphery soft tissue or subcutaneous space other cancer, with other cancer of the associated spleen cancer of lymph and leukemia or vascular system.Minimal medicament transport mechanism (brain, spinal column etc.) may also be beneficial to the effect of the cancer that is arranged in health, organ or privilege (privileged) biological space deeply.Particularly, the antitumor agent according to intradermal delivery of the present invention also can be used for treating pulmonary carcinoma.

The reinforced effects of intradermal delivery antitumor agent may be different for the medicament with different biological mechanism of action.The cellular toxicity medicine may show more initial tumor location and kill and wound before whole body distributes, thereby the intensifier target tissue kills and wounds and side effect reduced to minimum.Yet the cellular toxicity medicine must be that the tissue of protection medicine-feeding part avoids dead immediately or downright bad preparation after administration.Cytotoxic agents is wrapping to the benefit that can strengthen the intradermal administration in fugitive liposome, granule or other carrier of protecting surrounding tissue.Initial at tumor host response or stimulate downtrod medicine of replying because near the location of replying tumor will have potential bigger benefit.Influencing immune medicine is that chemotactic factor, cytokine and other immunostimulant (example of IL-12 is best example) might have unusual benefit via this delivery approach.The medicine (for example receptor of tumor specific antibody, target tumor surface marker and the mark that combines with the tumour-specific receptor) of the chemicals of target tumor has been mixed in use may enjoy largest benefit, because their target tumor cells all physically and chemically.This class medicine includes but not limited to therapeutic antibodies; carry the granule of cytotoxic agents or carry the other medicines of tomour specific targeting mark, thereby or combine with tumor and to start other medicament of attacking by inherent biological mechanism (for example cellullar immunologic response or reply by the antitumor of complement-mediated).

Method of the present invention also comprises with tumor cell uses antitumor agent, and the vaccine effect at following tumor challenge is provided thus.

Medicament deposition by improving institute's delivery of therapeutics agents, tissue biological's availability, begin effect and removing faster faster, the invention provides improving one's methods of treatment disease (for example cancer).The invention provides in order to treat the method for disease (particularly cancer) at least a therapeutic agent of administration, comprise with controllable rate, volume and pressure medicament is delivered to the ID zone of experimenter's skin, make medicament be deposited in the ID zone and and absorb by the lymph vascular system.

Can include but not limited to following by the cancer and the associated conditions of method and composition treatment of the present invention: leukemia, include but not limited to acute leukemia, acute lymphoblastic leukemia, acute myeloid leukemia, such as myeloblast, promyelocyte, Myelomonocyte, mononuclear cell, erythroleukemia leukemia and myelodysplastic syndrome, chronic leukemia is such as, but not limited to chronic myeloid (granulocytic) leukemia, chronic lymphocytic leukemia, hairy cell leukemia; Polycythemia vera; Lymphoma, sick such as, but not limited to He Jiejinshi (Hodgkin), Fei Hejiejinshi is sick; Multiple myeloma is such as, but not limited to smouldering multiple myeloma, nonsecreting type myeloma, osteosclerotic myeloma, Plasmacytic leukemia, solitary plasmacytoma and extramedullary plasmacytoma; Walden Si Telunshi (Waldenstr  m ' s) macroglobulinemia; The undetermined MG of meaning; Benign monoclonal gammopathy; Heavy chain disease; Bone and connective tissue sarcoma are such as, but not limited to osteosarcoma, os osseum sarcoma, chondrosarcoma, outstanding Yin Shi (Ewing ' s) sarcoma, pernicious giant cell tumor, the fibrosarcoma of bone, chordoma, periosteal sarcoma, soft tissue sarcoma, angiosarcoma, fibrosarcoma, Ka Boxi (Kaposi) sarcoma, leiomyosarcoma, liposarcoma, lymphangiosarcoma, schwannoma, rhabdomyosarcoma, synovial sarcoma; The cerebral tumor includes but not limited to glioma, astrocytoma, brain stem glioma, ependymoma, oligodendroglioma, non-neuroglial tumor (nonglial tumor), acoustic nerve neurilemmoma, craniopharyngioma, medulloblastoma, meningioma, pinealoma, pineocytoma, constitutional brain lymphoma; Breast carcinoma includes but not limited to adenocarcinoma, lobule (minicell) cancer, intraductal carcinoma, marrow sample breast carcinoma, mucin breast carcinoma (mucinous breast cancer), pipe breast carcinoma (tubular breast cancer), nipple breast carcinoma (papillary breast cancer), Pei Jiteshi (Paget) disease and inflammatory breast carcinoma; Adrenal carcinoma includes but not limited to pheochromocytoma and adrenocortical carcinoma; Thyroid carcinoma is such as, but not limited to the thyroid carcinoma (anaplasticthyroid cancer) of nipple or folliculus thyroid carcinoma (papillary or follicularthyroid cancer), medullary thyroid carcinoma and a change; The pancreas cancer includes but not limited to insulinoma, gastrinoma, glucagonoma, VIPoma (vipoma), Somat secreted tumor and carcinoid tumor or islet cell tumor; The hypophysis cancer includes but not limited to Ku Xinshi (Cushing ' s) disease, prolactin antagonist secreted tumor, acromegaly and diabetes insipius; Cancer eye includes but not limited to ophthalmomelanoma (such as iris melanoma, choroidal melanoma and cilliarybody melanoma) and retinoblastoma; Cancer of vagina includes but not limited to squamous cell carcinoma, adenocarcinoma and melanoma; Carcinoma vulvae includes but not limited to squamous cell carcinoma, melanoma, adenocarcinoma, basal cell carcinoma, sarcoma and Pei Jiteshi (Paget ' s) disease; Cervical cancer (cervicalcancer) includes but not limited to squamous cell carcinoma and adenocarcinoma; Uterus carcinoma (uterine cancer) includes but not limited to carcinoma of endometrium and sarcoma of uterus; Ovarian cancer includes but not limited to epithelial ovarian cancer, borderline tumor, blastoma and stromal tumor; Esophageal carcinoma includes but not limited to squamous cell carcinoma, adenocarcinoma, adenoid cystic carcinoma (adenoid cyctic carcinoma), mucoepidermoid carcinoma, adenosquamous carcinoma, sarcoma, melanoma, plasmocytoma, verrucous carcinoma and oat cells (minicell) cancer; Gastric cancer includes but not limited to adenocarcinoma, fungate (polypoid), ulcer, surperficial propagation, wide-scale distribution, malignant lymphoma, liposarcoma, fibrosarcoma and carcinosarcoma; Colon cancer; Rectal cancer; Hepatocarcinoma includes but not limited to hepatocarcinoma and hepatoblastoma (hepatoblastoma); Carcinoma of gallbladder includes but not limited to adenocarcinoma; Cancer of biliary duct includes but not limited to folliculus (pappillary), joint knot property and dispersivity; Pulmonary carcinoma includes but not limited to nonsmall-cell lung cancer, squamous cell carcinoma (epidermoid carcinoma), adenocarcinoma, large cell carcinoma and small cell lung cancer; Carcinoma of testis includes but not limited to (anaplastic) of germinoma, spermocytoma, a change, classical (typically), spermatocyte (spermatocytic), nonseminoma, embryonal carcinoma, teratoma, choriocarcinoma (yolk sac tumor); Carcinoma of prostate includes but not limited to adenocarcinoma, leiomyosarcoma and rhabdomyosarcoma; Penal cancers; Oral cancer includes but not limited to squamous cell carcinoma; The substrate cancer; Salivary-gland carcinoma includes but not limited to adenocarcinoma, mucoepidermoid carcinoma and adenoid cystic carcinoma (adenoidcystic carcinoma); Pharyngeal cancer includes but not limited to squamous cell carcinoma and wart; Skin carcinoma includes but not limited to basal cell carcinoma, squamous cell carcinoma and melanoma, surface propagation melanoma, nodular melanoma, lentigo maligna melanoma, acral-lentiginous melanoma; Renal carcinoma includes but not limited to renal cell carcinoma, adenocarcinoma, hypernephroma, fibrosarcoma, transitional cell carcinoma (renal pelvis and/or uterus uterer); Wei Ermusishi (Wilms ' s) tumor; Bladder cancer includes but not limited to transitional cell carcinoma, squamous cell carcinoma, adenocarcinoma, carcinosarcoma.In addition, cancer comprises that (summary of these diseases is seen Fishman etc. for myxosarcoma, osteogenic sarcoma, endotheliosarcoma, lymphangioendothelioma, mesothelioma, synovioma, hemangioblastoma, epithelial cancer, cystadenocarcinoma, bronchogenic carcinoma, syringocarcinoma, sebaceous gland carcinoma, papillary carcinoma and papillary adenocarcinoma, 1985, Medicine, the 2nd edition, J.B.Lippincott company, Philadelphia; With Murphy etc., 1997, Informed Decisions:TheComplete Book of Cancer Diagnosis, Trea tment, and Recovery, Viking Penguin, Penguin Books.U.S.A., Inc., the U.S.).

Therefore, method and composition of the present invention also can be used for treatment or prevents multiple cancer or other abnormality proliferation disease, include but not limited to: cancer comprises the cancer of bladder, breast, colon, kidney, liver, lung, ovary, pancreas, stomach, prostate, cervix uteri, thyroid and skin; Squamous cell carcinoma; The hemopoietic tumor of lymph pedigree comprises leukemia, acute lymphoblastic leukemia, acute lymphoblastic leukemia, B cell lymphoma, t cell lymphoma, Burketts lymphoma; The hemopoietic tumor of bone marrow pedigree comprises acute and chronic lymphocytic leukemia and promyelocyte leukemia; Between the tumor of matter origin, comprise fibrosarcoma and rhabdomyosarcoma; Other tumor comprises melanoma, spermocytoma, teratocarcinoma, neuroblastoma and glioma; The tumor of maincenter and peripheral nervous system comprises astrocytoma, neuroblastoma, glioma and schwannoma; Between the tumor of matter origin, comprise fibrosarcoma, rhabdomyosarcoma and osteosarcoma; And other tumor, comprise melanoma, xenoderma pegmentosum, keratoacanthoma (keratoactanthoma), spermocytoma, thyroid follcular carcinoma and teratocarcinoma.Also imagination also will be treated by method and composition of the present invention by the not normal cancer that causes of apoptosis.These cancers can include but not limited to the cancer of follicular lymphoma, p53 sudden change, breast, prostate and ovarian tumor and premalignant lesion such as the familial adenomatous polyposis and the myelodysplastic syndromes of dependence hormone.In specific embodiment, by pernicious in method and composition treatment of the present invention or prevention ovary, bladder, breast, colon, lung, skin, pancreas or the uterus or proliferative disorder variation (giving birth to and abnormal development) or excess proliferative disease such as changing.In other specific embodiments, by method and composition treatment of the present invention or prevention sarcoma, melanoma or leukemia.

The method that is used at experimenter's treatment or prophylaxis against infection diseases is also contained in the present invention, comprises that one or more medicaments with treatment or prevention effective dose are administered to the ID of experimenter's skin.Can be the infectious disease that causes by infective agents such as including but not limited to virus, antibacterial, fungus, protozoacide and virus by the infectious disease of molecular therapy of the present invention or prevention.

Can use the viral disease of method treatment of the present invention or prevention to include but not limited to by hepatitis A virus, hepatitis B virus, hepatitis C virus, influenza virus, chickenpox virus, adenovirus, I herpes simplex virus type (HSV-I), II herpes simplex virus type (HSV-II), rinderpest, rhinovirus, ECHO virus, rotavirus, respiratory syncytial virus, human papillomavirus, papovavirus, cytomegalovirus, echino virus (echinovirus), arbovirus, huntavirus, Coxsackie virus, mumps virus, Measles virus, rubella virus, poliovirus, variola, Epstein-Barr virus, I type human immunodeficiency virus (HIV-I), II type human immunodeficiency virus (HIV-II) and such as Viral miningitis, encephalitis, the disease that the material of virosis such as dengue fever or variola causes.

In some embodiment, the method that is used for the treatment of or prevents respiratory tract disease or disease (particularly respiratory tract infection, viral and bacillary) is contained in the present invention, comprises the intradermal zone that the therapeutic agent of effective dose is administered to the experimenter of needs treatment.The method of the respiratory tract infection of treatment or prevention upper respiratory tract (for example nose, ear, nasal sinuses and throat) and lower respiratory tract (for example trachea, bronchus and lung) is contained in the present invention.The example that causes the virus of upper respiratory tract infection comprises rhinovirus and influenza virus A and B.The example of lower respiratory channel virus infection has parainfluenza virus to infect (" PIV "), respiratory syncytial virus (" RSV ") and bronchiolitis.The example that causes the antibacterial of lower respiratory infection comprises streptococcus pneumoniae (Streptococcus pneumoniae) that causes pneumococcal pneumonia (pneumonococcalpneumonia) and the pulmonary tuberculosis mycobacteria (Mycobacterium tuberculosis) that causes tuberculosis.Comprise systemic candidiasis, blastomycosis, cryptococcosis (crytococcosis), coccidioidomycosis and aspergillosis by fungus-caused respiratory tract infection.Respiratory tract infection can be constitutional or secondary infection.

The invention provides and be used to prevent, control, treat or improve the respiratory tract disease that caused by any viral infection or relevant with it or the method for disease, described method comprises one or more therapeutic agents according to method effective dosage of the present invention.The viral example that causes viral infection includes but not limited to retrovirus (for example I and II type human T-cell lymphotrophic virus (humanT-cell lymphotrophic virus type I and II) (HTLV) and human immunodeficiency virus (HIV)), herpesvirus (for example I and II herpes simplex virus type (HSV), Epstein-Barr virus, HHV6-HHV8 and cytomegalovirus), arenavirus (for example Lassa fever virus), paramyxovirus (Measles virus for example, the human respiratory syncytial virus, mumps virus, hMPV and pneumonitis virus), adenovirus, Bunyavirus (bunyaviruses) (for example Hantaan virus), coronavirus, filamentous virus (filoviruses) (for example Ebola (Ebola) virus), banzi virus (hepatitis C virus (HCV) for example, yellow fever virus and Japanese encephalitis virus), hepadnavirus (hepadnavirus) (for example hepatitis B virus (HBV)), orthomyxovirus (orthomyoviruses) (influenza virus A for example, B, C and PIV), papovavirus (for example human papillomavirus), picornavirus (rhinovirus for example, enterovirus and hepatitis A virus), poxvirus, reovirus (for example rotavirus), togavirus (for example rubella virus) and rhabdovirus (for example rabies virus).At the biological answer-reply of viral infection include but not limited to that antibody horizontal raises, the propagation of T cell and/or soak into raises, the propagation of B cell and/or soak into rising, epithelial hyperplasia and generation mucin.The present invention also provides prevention, treatment, control or has improved the method for viral respiratory tract infection, such as virus (metapneumavirus) and adenopathy viral disease (for example heat generation respiratory tract disease, croup, bronchitis, pneumonia) after common cold, viral pharyngitis, viral laryngitis, viral croup, viral bronchitis, influenza, parainfluenza virus viral disease (" PIV ") (for example croup, bronchiolitis, bronchitis, pneumonia) and respiratory syncytial virus (" RSV "), the pneumonia, described method comprises one or more therapeutic agents of effective dosage.

In preferred embodiments, the method that is used for the treatment of disease or disease (particularly respiratory tract disease) is contained in the present invention, is included between the delivery of intranasal (IN) and whole body and splits the standard dose that is used for this sick therapeutic agent.Those skilled in the art can determine to split the best ratio of dosage by normal experiment.In the situation that IN/IM delivers, the ratio between preferred 5/95 and 30/70.The inventor finds that this causes the immediately level of therapeutic agent (for example antibody) in tissue (for example lung) consistent with concentration with infectious titer in external.In addition, split the preventive concentration that dosage can not cause keeping several weeks after administration detectable loss takes place.

Can use the method treatment of the invention relevant with the inventive method or the bacterial disease of prevention is by including but not limited to the mycobacterium rickettsia, mycoplasma, Neisseria, streptococcus pneumoniae, B. burgdorferi (Borrelia burgdorferi) (Lyme (Lyme) disease), Bacillus anthracis (Bacillus antracis) (anthrax), tetanus, streptococcus, staphylococcus, mycobacterium, tetanus, pertissus, cholera, the plague, diptheria, chlamydia, bacterial bacterial disease such as staphylococcus aureus and legionella.

Can use the method treatment or the prevention Protozoosis of the invention relevant with the inventive method is by the Protozoosis that includes but not limited to that Leishmania, kokzidioa, trypanosomicide or malaria protozoacidies such as (malaria) causes.

Can use the method treatment of the invention relevant with the inventive method or the parasitic disease of prevention is the parasitic disease that is caused by parasites such as including but not limited to chlamydia and rickettsia.

5.3 will be according to the medicament of administration of the present invention

The present invention is contained and is used for the treatment of, the biologic activity agent, particularly therapeutic agent of prevention or control disease or disease.The example that can be used for the biologic activity agent of the inventive method includes but not limited to immunoglobulin (polyspecific Ig for example, strand Ig, the Ig fragment), protein, peptide (peptide receptor for example, PNA, select albumen, conjugated protein (maltose-binding protein, glucose is conjugated protein)), nucleotide, nucleic acid (PNA for example, RNA, the RNA/DNA that modifies, fit (aptamers)), receptor (for example acetylcholinergic receptor), enzyme (glucoseoxidase for example, hiv protease and reverse transcriptase), carbohydrate (NCAM for example, sialic acid), cell (for example insulin and glucose responding sexual cell), phage (for example filobactivirus), virus (for example HIV), chemistry special dose (Chemospecific agents) (Cyptands for example, crown ether, Boronates).

The invention provides the method that is used for the administration antitumor agent.These antitumor agents comprise multiple reagent agent, comprise the anticarcinogen and the therapeutic antibodies of cytokine, angiogenesis inhibitor, classics.Can include but not limited to interferon, interleukin (IL-1 ,-2 ,-4 ,-6 ,-8 ,-12) and cell growth factor according to cytokine, immunoregulation agent and the hormone that the present invention uses.

The angiogenesis inhibitor that can be used for method and composition of the present invention includes but not limited to: angiostatin (plasminogen fragment); The Antithrombin III of angiogenesis inhibitor; Angiozyme; ABT-627; Bay 12-9566; Benfluralin (Benefin); Bevacizumab (Bevacizumab); BMS-275291; By the deutero-inhibitor of cartilage (CDI); CAI:CD59 complement fragment; CEP-7055; Col 3; Combretastatin A-4; Endostatin (collagen protein XVIII fragment); CH-296; Gro-β; Halofuginone; Heparinase; Heparin hexasaccharide fragment; HMV833; Human chorionic gonadotropin (hCG); IM-862; Interferon-ALPHA/β/γ; Interferon inducible protein matter (IP-10); Il-1 2; Kringle 5 (plasminogen fragment); Marimastat; Inhibitors of metalloproteinase (TIMP); The 2-methoxyestradiol; MMI 270 (CGS 27023A); MoAb IMC-1C11; Neovastat; NM-3; Panzem:PI-88; Placenta Hominis ribonucleic acid enzyme inhibitor; Inhibitors of plasminogen activator inhibitor; PF4 (PF4); Prinomastat; Prolactin antagonist 16kD fragment; Proliferin associated protein (PRP); PTK 787/ZK 222594; Biostearin; Solimastat; Squalamine; SS 3304; SU 5416; SU 6668; SU 11248; Tetrahydrocortisol-S; Tetrathiomolybdate; Neurosedyn; Thrombospondin-1 (Thrombospondin-1) (TSP-1); TNP-470; Transforming growth factor-beta (TGF-b); Angiostatin; Vasostatin (calreticulin fragment): ZD 6126; ZD 6474; Farnesyl transferase inhibitor (FTI); And diphosphonate (bisphosphonate).

Can include but not limited to according to other anticarcinogen that method of the present invention is used: acivicin (acivicin); Aklavine; The hydrochloric acid acodazole; Acronine; Adozelesin; Aldesleukin (aldesleukin); Altretamine; Ambomycin (ambomycin); The acetic acid Ametantrone; Aminoglutethimide; Amsacrine; Anastrozole (anastrozole); Anthramycin; Asparaginase; Asperlin; Azacytidine; Azetepa; Azotomycin; Batimastat; Dualar; Than mucositis amine (bicalutamide); The hydrochloric acid Bisantrene; Bisnafidedimesylate; Bizelesin; Bleomycin Sulphate; Brequinar sodium (brequinarsodium); Bropirimine; Busulfan; Actinomycin C; Calusterone (calusterone); Caracemide (caracemide); Carbetimer (carbetimer); Carboplatin; Carmustine; The hydrochloric acid carminomycin; Carzelesin (carzelesin); Cedefingol; Chlorambucil; Cirolemycin; Cisplatin; Cladribine; Crisnatol mesylate; Cyclophosphamide; Cytosine arabinoside; Dacarbazine; Actinomycin D; Daunorubicin hydrochloride; Decitabine (decitabine); Dexormaplatin; Dezaguanine; The methanesulfonic acid Dezaguanine; Diaziquone; Docetaxel (docetaxel); Amycin; Doxorubicin hydrochloride; Droloxifene; Droloxifene citrate; Dromostanolone propionate; Diazomycin; Edatrexate; Hydrochloric acid Chinese mugwort fluorine ornithine; Elsamitrucin; Enloplatin; Enpromate; Eponate; Epirubicin hydrochloride; Erbulozole; The hydrochloric acid Esorubicin; Estramustine; Estramustine phosphate sodium; Etanidazole (etanidazole); Etoposide; The phosphoric acid etoposide; Etoprine; Salt acid system sieve azoles azoles; Fazarabine (fazarabine); Fenretinide (fenretinide); Floxuridine; Fludarabine phosphate; Fluorouracil; Flurocitabine; Fosquidone (fosquidone); Fostriecin sodium (fostriecin sodium); Gemcitabine; Gemcitabine hydrochloride; Hydroxyurea; The hydrochloric acid darubicin; Ifosfamide; Ilmofosine; Interleukin (comprising recombinant interleukin 12 or rIL12); Intederon Alpha-2a; Interferon Alpha-2b; Interferon alfa-n1; Alferon N; Interferon beta-Ia; Interferon gamma-Ib; Iproplatin; Irinotecan hydrochloride (irinotecan hydrochloride); Lanreotide acetate (lanreotide acetate); Letrozole (letrozole); Leuprorelin acetate; Liarozole hydrochloride (liarozole hydrochloride); Lometrexol sodium; Lomustine; Losoxantrone hydrochloride; Aetinex; Maytansine; Mustine hydrochlcride; Megestrol acetate; Melengestrol acetate; Melphalan; Menogaril (menogaril); Mercaptopurine; Methotrexate; Methotrexate sodium; Metoprine; Meturedepa (meturedepa); Mitindomide (mitindomide); Mitocarcin; Mitochromine mitocromine B-35251; Mirincamycin; Mitomalcin; Mitomycin; Mitosper; Mitotane (mitotane); Mitoxantrone hydrochloride; Mycophenolic acid; Thiophene ammonia ester reaches azoles; Nogalamycin; Ormaplatin (ormaplatin); Oxisuran; Paclitaxel; Asparaginase; Peliomycin (peliomycin); Pentamustine; Peplomycin Sulfate; Perfostamide; Amedel; A-20968; The hydrochloric acid piroxantrone; Plicamycin (plicamycin); Plomestane (plomestane); Porfimer (porfimer sodium); Porfiromycin; Prednimustine; Procarbazine hydrochloride; Puromycin; Puromycin hydrochloride; Pyrazofurin; Isopentenyladenosine; Rogletimide; Safingol (safingol); The hydrochloric acid Safingol; Semustine; Simtrazene (simtrazene); Sparfosate sodium; Sparsomycin; Spirogermanium hydrochloride; Spiromustine; Spiroplatin; Streptonigrin; Streptozotocin; Sulofenur; Tallysomycin; Tecogalan sodium (tecogalan sodium); Tegafur; Teloxandrone hydrochloride; Temoporfin; Moor former times for the Buddhist nun; Teroxirone; Testolactone; ITG; Thioguanine; The match TEPA; Tiazofurine (tiazofurin); Tirapazamine; The citric acid Toremitene; Trestolone acetate; Phosphoric acid triciribine (triciribine phosphate); Trimetrexate; The Trimetrexate glucuronate; Triptorelin; Tubulozole hydrochloride; Uracil mustard; Uredepa; Vapreotide (vapreotide); Dimension is for Lip river sweet smell (verteporfin); Vinblastine sulfate; Vincristine sulfate; Vindesine; Vindesine sulfate; Sulphuric acid vinepidine (vinepidinesulfate); The sulphuric acid vinglycinate; The sulphuric acid vinleurosine; Tartaric acid dimension promise agent shore; The sulphuric acid vinrosidine; The sulphuric acid vinzolidine; R 83842; Zeniplatin; Neocarzinostatin; Zorubicin hydrochloride.Preferred other cancer therapy drug has 5-fluorouracil and formyl tetrahydrofolic acid.

Can comprise therapeutic antibodies according to other example of the antitumor agent of method administration of the present invention, include but not limited to ZENAPAX ^(daclizumab) (Roche Pharmaceuticals, Switzerland), it is to be used for the inhibitive ability of immunity Humanized anti-CD 25 monoclonal antibody that the prophylaxis of acute kidney allograft repels; PANOREX ^TM, it is the IgG2a antibody (Glaxo Wellcome/Centocor) of the anti-17-IA cell surface antigen in Mus source; BEC2, it is Mus source antiidiotype (GD3 epi-position) IgG antibody (ImClone System); IMC-C225, it is inosculating antibody EGFRIgG antibody (ImClone System); VITAXIN ^TM, it is that humanized anti-alpha V β 3 integrates plain antibody (Applied Molecular Evolution/MedImmune); Smart M195, it is Humanized CD 3-resisting 3IgG antibody (Protein Design Lab/Kanebo); LYMPHOCIDE ^TM, it is the anti-CD22IgG antibody of humanization (Immunomedics); ICM3, it is the anti-ICAM3 antibody of humanization (ICOS Pharm); IDEC-114, it is the anti-CD80 antibody of primatesization (IDEC Pharm/Mitsubishi); IDEC-131, it is humanization anti-CD40L antibodies (IDEC/Eisai); IDEC-151, it is primatesization anti-CD 4 antibodies (IDEC); IDEC-152, it is the anti-CD23 antibody of primatesization (IDEC/Seikagaku); The anti-CD3 of SMART, it is Humanized CD 3-resisting IgG antibody (Protein Design Lab); 5G1.1 it is the humanization anticomplementary factor 5 (C5) antibody (Alexion Pharm); D2E7, it is the anti-TNF-Alpha antibodies of humanization (CAT/BASF); CDP870, it is the anti-TNF-α of a humanization Fab fragment (Celltech); IDEC-151, it is the anti-CD4IgG1 antibody of primatesization (IDECPharm/SmithKline Beecham); MDX-CD4, it is the anti-CD4IgG antibody of people (Medarex/Eisai/Genmab); CDP571, it is the anti-TNF-α of a humanization IgG4 antibody (Celltech); LDP-02, it is humanized anti-alpha 4 β, 7 antibody (LeukoSite/Genentech); Or thoClone OKT4A, it is the anti-CD4IgG antibody of humanization (Or tho Biotech); ANTOVA ^TM, it is a humanization anti-CD 40 L IgG antibody (Biogen); ANTEGREN ^TM, it is the anti-VLA-4IgG antibody of humanization (Elan); And CAT-152, it is people's anti-TGF-beta ₂Antibody (Cambridge Ab Tech).

Can be used for particularly preferred biologic activity agent of the present invention is therapeutic antibodies.Monoclonal antibody is contained in the present invention, multi-specificity antibody, human antibody, Mus source antibody, humanized antibody, synthetic antibody, chimeric antibody, polyclonal antibody, camelization antibody (camelizedantibodies), strand Fv (scFv), single-chain antibody, the Fab fragment, F (ab ') fragment, with the continuous bispecific Fv (sdFv) of disulfide bond, the epi-position binding fragment of any therapeutic antibodies that internalization antibody (intrabodies) and antiidiotype (anti-Id) antibody (for example comprising anti-Id and anti-Id antibody at antibody of the present invention) and disclosed herein and this area are known.The therapeutic antibodies that the present invention is contained includes but not limited to HERCEPTIN ^(Trastuzumab) (Genentech, CA), it is the Humanized anti-HER 2 monoclonal antibody that is used for the treatment of the transitivity patients with mastocarcinoma; REOPRO ^(abciximab) (Centocor), it is the antibody that is used to prevent glycoprotein iib/iiia receptor on the antiplatelet that grumeleuse forms; ZENAPAX ^(daclizumab) (Roche Pharmaceuticals, Switzerland), it is to be used for the inhibitive ability of immunity Humanized anti-CD 25 monoclonal antibody that the prophylaxis of acute kidney allograft repels; PANOREX ^TM, it is the IgG2a antibody (GlaxoWellcome/Centocor) of the anti-17-IA cell surface antigen in Mus source; BEC2, it is Mus source antiidiotype (GD3 epi-position) IgG antibody (ImClone System); IMC-C225, it is an inosculating antibody EGFR IgG antibody (ImCloneSystem); VITAXIN ^TM, it is that humanized anti-alpha V β 3 integrates plain antibody (AppliedMolecular Evolution/MedImmune); Campath 1H/LDP-03, it is a humanized anti-CD 52 IgG1 antibody (Leukosite); Smart M195, it is Humanized CD 3-resisting 3IgG antibody (Protein Design Lab/Kanebo); RITUXAN ^TM, it be inosculating antibody CD20IgG1 antibody (IDEC Pharm/Genentech, Roche/Zettyaku); LYMPHOCIDE ^TM, it is the anti-CD22 IgG of a humanization antibody (Immunomedics); ICM3, it is the anti-ICAM3 antibody of humanization (ICOS Pharm); IDEC-114, it is the anti-CD80 antibody of primatesization (IDECPharm/Mitsubishi); ZEVALIN ^TM, it is a radiolabeled Mus source anti-CD 20 antibodies (IDEC/Schering AG); IDEC-131, it is humanization anti-CD40L antibodies (IDEC/Eisai); IDEC-151, it is primatesization anti-CD 4 antibodies (IDEC); IDEC-152, it is the anti-CD23 antibody of primatesization (IDEC/Seikagaku); The anti-CD3 of SMART, it is Humanized CD 3-resisting IgG antibody (Protein Design Lab); 5G1.1 it is the humanization anticomplementary factor 5 (C5) antibody (Alexion Pharm); D2E7, it is the anti-TNF-Alpha antibodies of humanization (CAT/BASF); CDP870, it is the anti-TNF-α of a humanization Fab fragment (Celltech); IDEC-151, it is the anti-CD4 IgG1 of a primatesization antibody (IDECPharm/SmithKline Beecham); MDX-CD4, it is a people source anti-CD4 IgG antibody (Medarex/Eisai/Genmab); CDP571, it is the anti-TNF-α of a humanization IgG4 antibody (Celltech); LDP-02, it is humanized anti-alpha 4 β, 7 antibody (LeukoSite/Genentech); OrthoClone OKT4A, it is the anti-CD4 IgG of a humanization antibody (Ortho Biotech); ANTOVA ^TM, it is a humanization anti-CD 40 L IgG antibody (Biogen); ANTEGREN ^TM, it is the anti-VLA-4 IgG of a humanization antibody (Elan); And CAT-152, it is people's anti-TGF-beta ₂Antibody (Cambridge Ab Tech).Those skilled in the art will understand, and the antibody that this paper announces can be used for prevention or postpone the outbreak or the progress of morbid state (for example cancer, tumor growth, cancerometastasis or infectious disease) in prevention or treatment.

In some embodiment, the present invention is contained the special antibody of respiratory pathogen (for example parainfluenza, influenza A, influenza B, chlamydia or adenovirus).In preferred embodiments, Mus source RSV specific antibody (RSV48), humanization palivizumab or its chimeric derivant are contained in the present invention fully.

Below table 1 listed other example of the antibody that can use according to the present invention.

Table 1: can be according to the monoclonal antibody that is used for treatment of cancer of the present invention's use

Company	Product	Disease	Effective object
				Abgenix AltaRex	ABX-EGF OvaRex BravaRex	Cancer ovarian cancer metastatic carcinoma	EGF receptor tumor antigen CA125 tumor antigen MUC1
Antisoma	Theragyn (pemtumomab 90Y) Therex	The ovarian cancer breast carcinoma	PEM antigen PEM antigen

Company	Product	Disease	Effective object
				Boehringer Ingelheim	Blvatuzumab	Head and neck cancer	CD44
Centocor/J&J Corixa	Panorex ReoPro ReoPro ReoPro Bexocar	Colorectal carcinoma PTCA acute MI ischemic stroke NHL	17-1A gp IIIb/IIIa gp IIIb/IIIa gp IIIb/IIIa CD20
				CRC Technology	MAb, antiidiotype 105AD7	The colorectum Theratope	gp72
Crucell	Anti-EpCAM	Cancer	Ep-CAM
				Cytoclonal	MAb, pulmonary carcinoma	Nonsmall-cell lung cancer	NA
Genentech	Herceptin Herceptin Rituxan Rituxan MAb-VEGF MAb-VEGF AMD Fab	Rudimentary or folliculus NHL middle rank and the senior NHL metastatic NSCLC metastatic colorectal cancer age related macular degeneration of metastatic breast cancer early-stage breast cancer recurrence/refractory	HER-2 HER-2 CD20 CD20 VEGF VEGF CD18
					E-26(2nd gen.IgE)	Allergic asthma and rhinitis	IgE
IDEC ImClone	Zevalin (Rituxan+ Yttrium-90) Cetuximab+innotecan Cetuximab+ cis-platinum and radiation Cetuximab+gemcitabine Cetuximab+ cis-platinum+5FU or Taxol Cetuximab+carboplatin+paclitaxel Cetuximab+ cis-platinum Cetuximab+ radiation BEC2+Bacillus Calmette Guerin BEC2+Bacillus Calmette Guerin IMC-1C11	The non-small cell lung cancer head of the head new diagnosis of colorectal cancer or repeatedly of the NHL refractory of the positive B cell NHL of the CD20 of rudimentary folliculus, relapsed or stubborn and Rituximab refractory and the metastatic pancreas cancer repeatability of the new diagnosis of neck cancer or metastatic head and the new diagnosis of neck cancer and the local senior head of neck cancer (widely incurable local disease and DISTANT METASTASES IN) and neck cancer ED-SCLC melanoma are with the colorectal cancer of hepatic metastases	CD20 EGF acceptor EGF acceptor EGF acceptor EGF acceptor EGF acceptor EGF acceptor EGF acceptor Ganglioside, GD3 analogies Ganglioside, GD3 analogies vegf receptor
				ImmonoGen	nuC242-DM1	Colorectum, stomach and pancreas cancer	nuC242

Company	Product	Disease	Effective object
				ImmunoMedics	LymphoCide LymphoCide Y-90 CEA-Cide CEA-Cide Y-90 CEA-Scan, (arcitumomab of Tc-99m mark) CEA-Scan, (arcitumomab of Tc-99m mark) CEA-Scan, (arcitumomab of Tc-99m mark) CEA-Scan, (arcitumomab of Tc-99m mark) LeukoScan, (sulesomab of Tc-99m mark) LymphoScan, (Tc-99m mark) AFP-Scan, (Tc-99m mark)	Tumour (radiophotography) soft tissue infection (radiophotography) lymthoma (radiophotography) liver 7gem-cell cancer (radiophotography) in non_hodgkin lymphoma non_hodgkin lymphoma metastatic solid tumors metastatic solid tumors colorectal cancer (radiophotography) breast cancer (radiophotography) lung cancer (radiophotography) operation	CD22 CD22 CEA CEA CEA CEA CEA CEA CEA CD22 AFP
Intracel	HumaRAD-HN (+90Y)	Head and neck cancer	NA
				Medarex MedImmune Merck KGaA	HumaSPECT MDX-101, (CTLA-4) MDX-210, (her-2 overexpression) MDX-210/MAK Vitaxin MAb 425 IS-IL-2	Colorectum imaging prostate and the various cancers of other various cancers of cancer prostate cancer cancer cancer	NA CTLA-4 HER-2 HER-2 αvβ 3EGF receptor Ep-CAM
Millennium NeoRx	Campath (alemtuzumab) CD20-Streptavidin (+biotin-yttrium 90) Avidicin (albumin+NRLU13)	Chronic lymphocytic leukemia non_hodgkin lymphoma metastatic carcinoma	CD52 CD20 NA
				Peregrine	Oncolym (+iodine-131) Cotara (+iodine-131)	The unresectable glioblastoma of non_hodgkin lymphoma	HLA-DR10 β DNA related protein
Pharmacia Corporation	C215 (+staphyloentero-toxin) MAb nacolomab tafenatox (C242+ staphyloentero-toxin)	Pancreas cancer lung and renal carcinoma colon and pancreas cancer	NA NA NA
				Protein Design	Nuvion	The T cell malignancies	CD3

Company	Product	Disease	Effective object
				Labs Titan	SMART M195 SMART 1D10 CEAVac TriGem TriAb	Senior colorectal carcinoma metastatic melanoma of AML NHL and small cell lung cancer transitivity breast carcinoma	CD33 HLA-DR antigens c EA GD2 ganglioside MUC-1
Trilex	CEAVac TriGem TriAb	Senior colorectal carcinoma metastatic melanoma and small cell lung cancer transitivity breast carcinoma	CEA GD2 ganglioside MUC-1
				Viventia Biotech	Radiolabeled NovoMAb-G2 Monopharm C GlioMAb-H (+gelonin)	Non_hodgkin lymphoma colorectum and pancreas cancer glioma, melanoma and neuroblastoma	NA SK-1 antigen NA
Xoma	Rituxan Rituxan ING-1	Recurrence/the rudimentary or folliculus NHL of refractory middle rank and senior NHL adenocarcinoma	CD20 CD20 Ep-CAM

The therapeutic agent that can be used for the present composition includes but not limited to chemotherapeutant, radiotherapy dose, hormone therapy agent, immunotherapeutic agent, immunoregulation agent, antiinflammatory, antibiotic, antiviral agent and cytotoxic agents.

The limiting examples of antiinflammatory comprises nonsteroidal and-inflammatory drug (NSAID), steroid anti-inflammatory drug, beta-2-agonists, anticholinergic agents (anticholingeric agent) and methylxanthine.The example of NSAID includes but not limited to aspirin, ibuprofen, celecoxib (celecoxib) (CELEBREX ^TM), diclofenac (VOLTAREN ^TM), etodolac (LODINE ^TM), Fei Nuoluofen (NALFON ^TM), indomethacin (INDOCIN ^TM), ketorolac (ketoralac) (TORADOL ^TM),  promazine (DAYPRO ^TM), nabumetone (RELAFEN ^TM), sulindac (CLINORIL ^TM), tolmetin (TOLECTIN ^TM), rofecoxib (rofecoxib) (VIOXX ^TM), naproxen (ALEVE ^TM, NAPROSYN ^TM), ketoprofen (ACTRON ^TM) and nabumetone (RELAFEN ^TM).These NSAID bring into play function by suppressing cyclo-oxygenase (for example COX-1 and/or COX-2).The example of steroid anti-inflammatory drug includes but not limited to glucocorticoid, dexamethasone (DECADRON ^TM), cortisone, hydrocortisone, prednisone (DELTASONE ^TM), prednisolone, triamcinolone, sulfasalazine and eicosanoid (such as prostaglandin, thromboxane and leukotriene).

The example of immunoregulation agent includes but not limited to methotrexate, ENBREL, REMICADE ^TMLeflunomide, cyclophosphamide, Ciclosporin A, and macrolide antibiotic (for example FK506 (fujimycin 506)), methylprednisolone (MP), 17-hydroxy-11-dehydrocorticosterone, steroid, mycophenolatemofetil, rapamycin (sirolimus), mizoribine, deoxyspergualin, brequinar, malononitriloamindes (for example leflunamide), TXi Baoshouti modulator and cytokine receptor modulator, 17-hydroxy-11-dehydrocorticosterone, the cytokine agonist, cytokine antagonist and cytokine inhibitor.

Antibiotic example includes but not limited to macrolide (tobramycin (Tobi for example ^)), cephalosporin (Cefalexin (Keflex for example ^), cefradine (Velosef ^), cefuroxime (Ceftin ^), cefprozil (cefprozil) (Cefzil ^), cefaclor (Ceclor ^), cefixime (Suprax ^) or cefadroxil (Duricef ^)), clarithromycin (clarithromycin (Biaxin for example ^)), erythromycin (erythromycin (EMycin for example ^)), penicillin (penicillin V (V-Cillin K for example ^Or Pen Vee K ^)) or quinolone (ofloxacin (Floxin for example ^), ciprofloxacin (Cipro ^) or norfloxacin (Noroxin ^)); aminoglycoside antibiotics (apramycin for example; arbekacin; bambermycin; Ambutyrosin.; dibekacin; neomycin; undecylenate; netilmicin sulfate; paromomycin; ribostamycin; sisomicin and Togoplus); amphenicol antibiotic (azidamfenicol for example; chloromycetin; florfenicol and thiamphenicol); ansamycin antibiotic (for example rifamide and rifampicin); carbacephems (for example Lorabid); carbapenems (for example biapenem and imipenum); cephalosporins (cefaclor for example; cefadroxil; cefadole; cefatrizine; cefazedone; cefozopran; cefpimizole; cefpiramide and cefpirome); cephamycin class (cefbuperazone for example; cefmetazole and cefminox); monocycle amide bacteriums (aztreonam for example; carumonam and tigemonam); oxacephems (for example Flomoxef and latamoxef); penicillins (nitrogen  amidine penicillin for example; volt nitrogen  amidine penicillin; the amoxicillin; bacampicillin; benzylpcnicillin acid; sodium benzylpenicillin; epicillin; Fenbenicillin; the flucloxacillin; penamecillin; the diethylaminoethylpenicillin G hydriodide; benethamine penicillin; penicillin 0; penicillin V; penicillin V benzathine; penicillin V hydrabamine; penimepicycline and phencihicillin potassium); Lincoln's amide-type (for example clindomycin and lincomycin); amphomycin; bacitracin; capreomycin; colistin; enduracidin; enviomycin; Tetracyclines (apicycline for example; duomycin; clomocycline and demeclocycline); 2,4-di-amino-pyrimidine (for example brodimoprim); itrofurans (for example furaltadone and furazolium chloride); quinolones and analog thereof (cinoxacin for example; clinafloxacin; flumequine and grepagloxacin); sulfonamides (sulfacetamide methoxy pyrazine for example; benzylsulfamide; noprylsulfamide; phthaloylsulfacetamide; prontosil and renoquid); sulfone class (thymolsufone for example; angeli's sulfone and solapsone); cycloserine; mupirocin; chloromycetin; erythromycin; penicillin; streptomycin; vancomycin; trimethoprim-sulfamethoxazolum azoles and tuberin.

The example of antiviral agent includes but not limited to protease inhibitor, the nucleoside reverse transcriptase mortifier, non-nucleoside reverse transcriptase mortifier and nucleoside analog, azidothymidine AZT, acyclovir, gangcyclovir, vidarabine, idoxuridine, trifluridine and ribavirin and phosphorus formic acid (foscarnet), amantadine, rimantadine, Saquinavir, indinavir, amprenavir, sieve that Wei, ritonavir, alpha-interferon, adefovirdipivoxil (adefovir), clevadine, Entecavir (entecavir) and pleconaril, ribavirin, rimantadine, amantadine, neuraminic acid enzyme inhibitor and many lipid drug derivatives, natural acid, phospholipid or docosahexenoic acid (DHA).

Can be used for other therapeutic agent of the present invention and include but not limited to α-1 antitrypsin, antiangiogenic agent, antisense, butorphanol, calcitonin and analog, Ceredase, the COX-II mortifier, the dermatological medicament, dihydroergotamine, dopamine agonist and antagonist, Enkephalins and other opioid peptides, epidermal growth factor, erythropoietin and analog, follicle stimulating hormone, G-CSF, glucagon, GM-CSF, granisetron, growth hormone and analog (comprising growth hormone releasing hormone), growth hormone antagonist, hirudin and hirudin analog are such as HIRULOG, IgE contains thing, insulin, insulinotropin and analog, insulin like growth factor, interferon, interleukin, lutropin, luteinizing hormone releasing hormone and analog, heparin, low molecular weight heparin is natural with other, that modify or synthetic glucosaminoglycan, M-CSF, metoclopramide, midazolam, monoclonal antibody, PEGization antibody, PEGization protein or any protein of modifying with hydrophilic or hydrophobic polymer or other functional group, fusion rotein, single chain antibody fragments or it and attachment protein matter, macromolecule, or any combination of other functional group, narcosis analgesic, nicotine, the non-steroid antiinflammatory, oligosaccharide, ondansetron, parathyroid hormone and analog, pth antagonist, prostaglandin antagonists, prostaglandins, the recombinant soluble receptor, scopolamine, combination of serotonin agonist and antagonist, Sildenafil, terbutaline, thrombolytics, tissue-type plasminogen activator, TNF and TNF antagonist, contain or do not contain the vaccine of carrier/adjuvant, comprise that preventive and therapeutic antigen (include but not limited to protein subunit matter, peptide, and polysaccharide, polysaccharide conjugates, toxoid, vaccine based on gene, the live body attenuation, reprovision, deactivation, full cell, virus and bacteria carrier), it is with following relevant: addiction, arthritis, cholera, cocaine addiction, diphtheria, tetanus, HIB, Lyme disease, meningococcus, measles, parotitis, rubella, chickenpox, yellow fever, respiratory syncytial virus, Ticks is propagated Japanese encephalitis, streptococcus pneumoniae, streptococcus, typhoid fever, influenza, hepatitis (comprises the first type, B-mode, third type, and hepatitis E), otitis media, rabies, poliomyelitis, HIV, parainfluenza, rotavirus, epstein-barr virus, CMV, chlamydia, atypia haemophilus (non-typeable haemophilus), moraxellacatarrhalis, the human papillomavirus, tuberculosis (comprising BCG), gonorrhea (gonorrhoea), asthma, atherosclerosis, malaria, escherichia coli, Alzheimers (Alzheimer) disease, helicobacter pylori, Salmonella, diabetes, cancer, herpes simplex, human papilloma etc., other material that comprises all main therapeutic agents is such as the medicament that is used for common cold, anti-addictive drug, antiallergic agent, the anti-emetic, appetrol (anti-obesity), anti-osteoporotic, anti-infective, analgesic, anesthetis, anoretics, anti-arthritic, antiasthmatics, anticonvulsant, antidepressants, antidiabetic drug, antihistaminic, the antibiotic medicine, the migraine preparation, anti-dizzy medicine, antinauseant, antitumor agent, antiparkinsonian drug, pruritus, psychosis, antipyretic, anticholinergic, the benzodiazepine derivatives antagonist, vasodilation (comprises comprehensively, coronarius, on every side with brain), the bone stimulant, central nervous system stimulant, hormone, sleeping pill, immunosuppressant, the muscular flaccidity agent, parasympatholytic, parasympathomimetic agent, prostaglandin, protein, peptide, polypeptide and other macromolecule, psychostimulant, tranquilizer, with sexual hypofunction and tranquilizer.

5.3.1 compositions

The compositions (or preparation) that comprises one or more biologic activity agent (particularly therapeutic agent) with solution form, particle form or its mixture is contained in the present invention.Employed compositions can be obtained by any species in the inventive method, perhaps generates by any recombinant DNA technology that those skilled in the art will know that.The compositions that comprises one or more biologic activity agent can include but not limited to pig, cattle, sheep, horse etc. from different animal species.Can modify the chemical state of these medicaments by the recombinant DNA technology of standard, thereby generate the medicament of the different chemical prescription of different bonding states.

Wait to deliver or the form of the biologic activity agent of administration comprises that solution, emulsion, suspension, gel, granule (such as miniature and nano-particle, or suspend or disperse) and original position that they can be accepted in diluent or the solvent at pharmacopedics form carrier.Compositions of the present invention can take to be suitable for any form of intradermal delivery.In one embodiment, intradermal compositions of the present invention is taked the form of mobile injectable medium, i.e. the low viscosity compositions that can inject in syringe or pen.Mobile injectable medium can be a liquid.Perhaps, mobile injectable medium is the liquid of particulate matter of wherein having suspended, and makes medium keep its flowability, thereby can inject and inject, for example can administration in syringe.

In some embodiment, preparation of the present invention comprises medicament and one or more other additives for the treatment of effective dose.The additive that can be used for preparation of the present invention comprises for example material or the pH buffer agent of wetting agent, emulsifying agent, change insulin quarternary structure.Preparation of the present invention can comprise one or more other excipient, such as sugar and polyhydric alcohol.Pharmacopedics can be accepted other example of carrier, diluent and other excipient and see Remington ' s PharmaceuticalSciences, Mack publishing company, N.J., current version (all complete being collected herein by reference).

Wait to deliver or the form of the therapeutic agent of administration comprises that solution, emulsion, suspension, gel, granule (such as miniature and nano-particle, or suspend or disperse) and original position that they can be accepted in diluent or the solvent at pharmacopedics form carrier matter.Preparation of the present invention can take to be suitable for any form of intradermal delivery.In one embodiment, intradermal preparation of the present invention is taked the form of mobile injectable medium, i.e. the low viscosity preparation that can inject in syringe or novopen.Mobile injectable medium can be a liquid.Perhaps, mobile injectable medium is the liquid of particulate matter of wherein having suspended, and makes medium keep its flowability, thereby can inject and inject, for example can administration in syringe.

Intradermal preparation of the present invention can be prepared into unit dosage form.Every bottle of unit dose can contain the 0.1-0.5ml preparation.In some embodiment, the unit dosage form of intradermal preparation of the present invention can contain 50 μ l-100 μ l, 50 μ l-200 μ l or 50 μ l-500 μ l.If desired, can these preparations be adjusted to expectation concentration by in each medicine bottle, adding sterile diluent.According to the preparation of the inventive method administration in a large number (in volumes) use, otherwise intradermal space may overload, and causes being assigned to one or more other zones, such as the SC zone.

Employed therapeutic agent can be liquid or powder type in the inventive method.In specific embodiment, when the intranasal administration preparation, liquid form will comprise therapeutic agent (for example antibody), will be with drop or atomised form administration.

The powder type of therapeutic agent will comprise the powder of the several different methods preparation of knowing by this area, and for example lyophilizing, spray are done or sprayed lyophilization (SFD) method, for example described in the U. S. application number 10/299,012 (with its complete being collected herein by reference).

The powder type of medicament can comprise therapeutic agent purely, perhaps can comprise one or more other compositions.Generally speaking, can use multiple conventional liq that the therapeutic interest agent is mixed with liquid preparation at first.Preferably, liquid is the liquid of aqueous, such as for example (for example injectable grade water) or multiple conventional buffer (contain or not saliferous).Buffer is selected to make the protein of selection or the therapeutic agent stable p H of other type usually, and normally those skilled in the art are confirmable.Generally speaking, this is in physiology pH scope, although some protein also is stable in wideer pH scope, and acid pH for example.Thus, the preferred pH scope of original liquid preparation is that about 1-is about 10, and especially preferably about 3-is about 8, especially preferably about 5-about 7.Those skilled in the art will understand, and can adopt extremely multiple suitable buffer.Suitable buffer includes but not limited to sodium acetate, sodium citrate, sodium succinate, ammonium bicarbonate sodium and sodium carbonate buffer.Generally speaking, using the molar concentration of buffer agent is the about 2M of about 1mM-, the about 1M of preferably about 2mM-, the about 0.5M of especially preferably about 10mM-, preferred especially 50-200mM.Generally speaking, if there is salt in the liquid solution, the molar concentration of used salt is the about 2M of about 1mM-so, the about 1M of preferably about 2mM-, the about 0.5M of especially preferably about 10mM-, preferred especially 50-200mM.Suitable salt includes but not limited to NaCl.

Liquid preparation can be taked various ways, for example solution, suspension, emulsion (such as oil/water or water/oil/aqueous emulsion), slurry or colloid.

Optional is that liquid preparation can comprise one or more conventional pharmacopedicss can accept excipient." excipient " is often referred to chemical compound or the material that adds for the effect of the preparation of enhanced activity ingredient (API).Example for example comprise in order to ensure or improve stability and after this long-time stability and mobile and cryoprotective agent (cryoprotectants) and the lyoprotectants that add of powder-product of protein in spraying freezing dry process or in spraying freezing air drying process.Suitable protective agent is free flowing granule solid comparatively normally; not can with thicken or polymerization after water contacts; basically harmless after being sucked the back by the patient or otherwise entering in patient's body, and can in the mode that changes its biologic activity remarkable interaction not take place with therapeutic agent.Suitable excipient includes but not limited to protein, such as people and bovine serum albumin, gelatin, immunoglobulin; Carbohydrate comprises monosaccharide (for example galactose, D-mannose, sorbose etc.), disaccharide (for example lactose, trehalose, sucrose etc.), cyclodextrin and polysaccharide (for example Raffinose, maltodextrin, glucosan etc.); Aminoacid, such as monosodium glutamate, glycine, alanine, arginine or histidine, and hydrophobic amino acid, for example tryptophan, tyrosine, leucine, phenylalanine etc.; Methylamine is such as betanin; Excipient salt is such as magnesium sulfate; Polyhydric alcohol, such as trihydroxy or senior sugar alcohol, for example glycerol (glycerin), erythritol, glycerol (glycerol), 1,2,3,4,5-pentanepentol, xylitol, Sorbitol and mannitol; Propylene glycol; Polyethylene Glycol; Pluronics; Surfactant; And combination.Preferred excipient comprises for example trehalose, sucrose and mannitol.Usually using another kind of excipient is that mucoadhesive (mucoadhesive) increases contacting of API and mucomembranous surface.The example of mucoadhesive comprises for example chitosan, dermatan sulfate, chrondroitin and pectin.In addition, can in the liquid preparation of the SFD method that is suitable for this paper announcement, add the deliquescent conventional cosolvent of improvement API.

Generally speaking, when using mucoadhesive, their amount ranges is about 1-95wt%, preferably about 1-50wt%, especially preferably about 5-50wt%, especially preferably about 5-20%.Generally speaking, the working concentration of cryoprotective agent is between about 5wt% and about 95wt%.

Dry powder of the present invention can be mixed with filler (bulking agent) or carrier, be used for reducing the concentration of therapeutic agent at the powder that the patient is delivered; The material volume that promptly may wish each unit dose is bigger.Filler can also be used to improve the performance characteristic of powder.Normally crystallization of suitable filler (to avoid the absorption of water) includes but not limited to lactose and mannitol.Therefore, if add such as filleies such as lactose, the ratio of adding purpose therapeutic agent and filler can change, preferably by about 99: 1 to about 1: 99, and more preferably by about 1: 5 to about 5: 1, especially preferred about 1: 10 to about 1: 20.

The present invention is contained the intradermal administration compositions of announcing as this paper of the present invention and is united other delivery approach, comprises in for example subcutaneous-intradermal interface, intranasal (IN), parenteral (for example intramuscular, intraperitoneal, intravenous and subcutaneous), epidural and mucosa (for example intranasal and path, oral cavity), the tumor, tumor is outer, part and epidermis.Can use compositions by any conventional route, for example by infusion or inject, the absorption (for example oral mucosa, rectum and intestinal mucosa etc.) by epithelium or mucocutaneous lining, and can be with other biologic activity agent administration.Administration can be whole body or partial.In addition, can also adopt pulmonary administration, for example by use inhaler or aerosol apparatus, and the preparation that contains propellant.See for example U.S. Patent number 6,019,968; 5,985,320; 5,985,309; 5,934,272; 5,874,064; 5,855,913; 5,290,540; With 4,880,078; And PCT publication No. WO92/19244; WO97/32572; WO97/44013; WO98/31346; And WO99/66903 (all complete being collected herein by reference of each piece of writing).

5.3.2 the evaluation of treatment effectiveness

Preferably be used for human before external, the cell culture system and in animal model organism (such as the rodent models system) to aspect several of the desired therapeutic active testing present composition, preventive or therapeutic agent.For example, can be used for need determining whether the algoscopy of administration particular composition to comprise the cell culture algoscopy, wherein cultivate patient's tissue sample, expose or otherwise contact pharmacopedics compositions of the present invention, and observe of the effect of this compositions tissue sample.Tissue sample can be gathered by the patient by biopsy.This test can be identified the most effective preventative or therapeutic molecules in the treatment for every individual patients.In a plurality of specific embodiments, can use the representative cell or the cell type (for example T cell) that relate to autoimmune or inflammatory disease to carry out the external test method, to determine whether pharmaceutical composition of the present invention has desired effects to these cell types.

Can be in the suitable animal model system test preventive and/or the combination of therapeutic agent before being used for the mankind.These animal model systems include but not limited to rat, mice, chicken, cattle, monkey, pig, Canis familiaris L., rabbit etc.Can adopt any animal model well-known in the art.In a specific embodiments of the present invention, in mouse model system, tested the combination of preventive and/or therapeutic agent.The proper model system is used widely, and is well-known to those skilled in the art.Preventive and/or therapeutic agent can repeat administrations.Several aspects of flow process can change.Described aspect comprises the time scheme of administration preventive and/or therapeutic agent, and is separately or as mixture to use these reagent.

Can in cell culture or laboratory animal, measure the toxicity and the effect of the preventative and/or therapeutic scheme of the present invention by the standard pharmaceutical program, for example measure LD ₅₀(50% colony's fatal dose) and ED ₅₀(50% mass treatment effective dose).Dosage rate between murder by poisoning and the therapeutic effect is a therapeutic index, and it can be expressed as ratio LD ₅₀/ ED ₅₀Preferably have exponential preventive of bigger treatment and/or therapeutic agent.Although can use preventive and/or therapeutic agent with toxic side effect, however must careful design with the delivery system at these medicament targeting illing tissue positions, thereby will reduce to minimumly to the latent lesion of uninfection cell, reduce side effect thus.

The data that obtained by cell culture test and zooscopy can be used for formulating preventive and/or the dosage of therapeutic agent scope that is used for the mankind.The dosage of these medicaments preferably is in and comprises ED ₅₀And do not have toxicity or have only in a little toxic circulation composition scope.Dosage can change in this scope with the route of administration that is adopted according to the dosage form that is adopted.For employed any medicament in the inventive method, at first can be by cell culture test estimation treatment effective dose.Can formulate the dosage that reaches the circulating plasma concentration range in animal model, described scope is included in the IC that measures in the cell culture ₅₀(concentration when promptly the chemical compound of surveying reaches and suppresses the half value of symptom is maximum).These information can be used for being determined at more accurately dosage useful in the human body.Can measure level in the blood plasma by for example high performance liquid chroma-tography.

Can also use the kinds of experiments animal model that is used to study cancer to measure the active anticancer of the therapy of using according to the present invention, transgenic mice or nude mice such as SCID mouse model or carrier's xenograft, know such as this areas such as hamster, rabbits with Relevance of TumorModels for Anticancer Drug Development, 1999, Fiebig and Burger compile; Contributions to Oncology, 1999, Karger; The Nude Mouse inOncology Research, 1991, Boven and Winograd compile; And Anticancer DrugDevelopment Guide, 1997, Teicher compiles the animal model of describing in (complete being collected herein by reference).

Can use the cell of tumor or malignant clone to screen therapeutic agent and method.Many standard test methods of this area can be used for assessing these survivals and/or growing state; For example can be by measuring ³The H-thymidine mixes, directly cell counting, detect known (such as proto-oncogene, fos for example, myc) or the changes in mRNA transcription level of cell cycle mark measure cell proliferation; Can dye by trypan blue and assess the survival ability of cell; Can slow down and/or in soft agar, form colony or in three-dimensional substrates film or extracellular matrix prepared product, form the tubulose network by range estimation morphological change, growth and assess the differentiation situation; Or the like.

In addition, can use any algoscopy that those skilled in the art will know that to assess that conjoint therapy disclosed herein is used for the treatment of or the preventative and/or therapeutic effect of anti-cancer, inflammatory disease or autoimmune disease in advance.

6. embodiment

6.1 the delivery approach is to the influence (using 100ng dosage) of IL-12 anti-tumor activity

6.1.1 flow process

Give C57BL/6J mice (Charles Rivers Labs company) in right front rib SC inoculation 1 * 10 ⁶Individual B16F10 melanoma cells (ATCC).A week behind the inoculated tumour (the 7th day), with the mice randomization, and with the 100ng rmIL-12 (R﹠amp that is dissolved in 50 μ l PBS solution; D Systems) treat, by or the tumor inoculation position near ID or SC injection, or IP injection (n=25/ condition).The extra condition of ID contrast uses 50 μ l PBS solution to carry out separately.ID treatment is to use 34G, and the 1mm pin is according to amended Mang Tufa administration.SC and IP administration are to use standard 25G * 3/4, and " pin is implemented.Treatment was carried out every other day until the 13rd day, added up to 4 doses.7th, gathered measurement of tumor result, the mm of unit in 11,14,18,21,25 and 28 days ²(length x width).0th, 14,21 and 28 days blood sample collections are used for IL-12 and IFN-γ analysis.The the 14th and 24 day each condition got 5 animals and gathered the facs analysis that draining lymph nodes and spleen tissue sample are used for CD49b (NK) cell.Each condition animal previously selected by 10, that be not used for blood collection or facs analysis is gathered the mortality rate data.Mortality rate is greater than 400mm by the tumor size ²Natural death that causes or euthanasia are measured.Use the t check: Two-Sample AssumingUnequal Variances computational statistics data.See for example Brunda, M., 1993, J.Exp.Med.178:1223-1230; With Leonard etc., 1997, Blood 90:2541-2548 (all being collected herein by reference).

6.1.2FACS analyze

Each treatment group is got 5 mice excision right lateral surface groin draining lymph nodes (DLN) and spleen, DLN placed cold HBSS (Invitrogen Life Technologies is housed, Carlsbad in culture dish CA), and places cold erythrocyte splitting buffer (the 0.16M NH of 10ml with spleen ₄Cl and 10mM KHCO ₃, Sigma, St.Louis, MO) in.By mechanical damage every part of DLN and spleen are processed into single-cell suspension liquid.1: 20 diluent to the cell solution that obtains thus carries out cell counting.Cell is centrifugal 15 minutes in 4 ℃ with 1500rpm.The sucking-off supernatant with cell with 5ml HBSS buffer solution for cleaning once, and uses the same terms recentrifuge.The sucking-off supernatant, and with cell with 2-4 * 10 ⁸The density of individual cell/ml is resuspended in Pharmingen dyeing buffer, and (San Jose CA) is used for streaming dyeing for Pharmingen, BD Biosciences.With about 1 * 10 ⁷Individual re-suspended cell (25 μ l) is added in the hole of 96 orifice plates.The cell of Xiang Kongzhong adds dyeing mixed liquor (25 μ l) and passes through the piping and druming mixing.As required, mixed liquor contains the combination (every kind of concentration in Pharmingen dyeing buffer is 0.01mg/ml) of following traget antibody: FITC-CD49b (Pan-NK cell, clone DX5, Pharmingen, BD Biosciences, San Jose, CA); PE-CD19 (Pan B cell, clone 1D3, Pharmingen, BD Biosciences, San Jose, CA); CY5PE-B220 (granulocyte and mononuclear cell, clone RA3-6B2, Pharmingen, BD Biosciences, San Jose, CA); APC-CD4 (t helper cell, clone RM4-5, Pharmingen, BDBiosciences, San Jose, CA); And APC-CY7-CD8 (the T cytotoxic cell, clone BC-CD8a, Biocarta, San Diego, CA).Cell/dye mixture is incubated 1 hour in 4 ℃ in the dark.With 150 μ l FacsFlow buffer (Pharmingen, BDBiosciences, San Jose, CA) clean-out opening, and centrifugal 5 minutes in 4 ℃ with 1500rpm.The sucking-off supernatant, and clean once more.To be resuspended in the cold FacsFlow buffer of 1ml through the cell that cleans, and place in the dark on ice until using FACS Vantage SE to carry out flow cytometry.The designing and arranging of cell analysis is except the B cell, and quantification CD4+ cell, CD8+ cell and CD49b+ (NK) cell.See for example Cordaro, T., 2002, J.Immunology 168:651-660; Fogler etc., 1998, J.Immonology 161:6014-6021; Gao etc., 2003, J.Exp.Med.198 (3): 433-442; Harada etc., 1998, Int.J.Cancer 75:400-405; Jenne etc., 2000, Cancer Research 60:4446-4452; And Pa rk etc., 2003, J.Immunology 170:1197-1201 (all being collected herein by reference).

6.1.3 tumor growth

As shown in Figure 1, the IL-12 that has only ID to deliver shows suppressing the remarkable effect of tumor growth.In ID administration condition, saw in the 18th day be better than tumor size that IP delivers significantly (p＜0.0002) dwindle.Also seen and be better than the similar effect that ID delivers PBS (p＜0.0000005) and SC (p＜0.00003) condition.Significant trend continues to research and finishes promptly the 28th day.

IL-12 dosage in this research than the low 2-10 of effective IP dosage of announcing in the document doubly.See for example Tannenbaum etc., 1996, J.Immunology 156:693-699; With Tsung etc., 1997, J.Immunology 158:3359-3365.This may be exactly the reason that IP takes the response of IL-12 shortage in this research.This has shown that being better than other dosage of having delivered Therapeutic Method saves effect.Directly the targeting lymph will mean the reduction expense than low dosage, and reduce side effect probably.

6.1.4 mortality rate

As shown in Figure 2, the 28th day, the mortality rate of delivering the group (10 merely hit 7 survives) of accepting IL-12 by ID was lower than other group of accepting IL-12 by IP (10 merely hit 3 survives) or SC (10 merely hit 3 survives) delivery.And the mortality rate of delivering the group of accepting IL-12 by ID also is lower than the group of only accepting PBS by ID delivery (10 merely hit 3 survives).Thus, ID delivery IL-12 compares big by 200% in the survival rate of demonstration in the 28th day with delivery or contrast by other approach.

6.1.5NK cell counting

IFN-γ thinks the curative effect effect that IL-12 delivers.Verified, IL-12 increases the IFN-γ that is generated by NK and T cell, and promotes the propagation and the differentiation of NK cell.Thus, the effect of NK cytosis indication IL-12 therapy strengthens.Be not limited to concrete theory, the NK cytosis can be caused by following: NK cell proliferation or differentiation strengthen; IFN-γ generates and discharges and strengthens; Targeting to the tumor proliferation position is better; The immunoregulation process absorbs the better effects if of the efficient targeting of approach via lymph; And/or above-mentioned every combination.

As shown in Figure 3, behind a day of IL-12 therapeutic scheme (the 14th day), the increase of the NK cell of ID delivery group obviously is better than other condition, and in ID group relative reduction is arranged at the 24th day, but still shows and be better than other condition the 24th day relative increase.

6.1.6 statistical data

The following statistical data that is obtained by the 18th and 28 day t check has shown that ID delivers IL-12 and is better than the significance that all other tests are delivered.

The 18th day T testing data of table 1

IP is to ID
				IP 100ng	ID 100ng
Meansigma methods	158.2458	62.28816
			Variance (variance)	8780.02	2255.463
Observed value	19	19
			Assumed average is poor	0
Df	27
			The t statistics	3.981629
P(T＜＝t)one-tail	0.000232
			t Critical one-tail	1.703288
P(T＜＝t)two-tail	0.000465
			t Critical two-tail	2.051829
ID is to SC
				ID 100ng	SC 100ng
Meansigma methods	62.288163	199.7379
			Variance	2255.4628	12576.68
Observed value	19	18
			Assumed average is poor	0
Df	23
			The t statistics	-4.807544
P(T＜＝t)one-tail	3.761E-05
			t Critical one-tail	1.71387
P(T＜＝t)two-tail	7.523E-05
			t Critical two-tail	2.0686548
ID is to contrast
				ID 100ng	ID-PBS
Meansigma methods	62.288163	122.8243
			Variance	2255.4628	3168.4
Observed value	19	19
			Assumed average is poor	0

Df	35
			The t statistics	-3.582918
P(T＜＝t)one-tail	0.0005116
			t Critical one-tail	1.6895729
P(T＜＝t)two-tail	0.0010233
			t Critical two-tail	2.0301104

The 28th day t testing data of table 2

IP is to ID
				IP 100ng	ID 100ng
Meansigma methods	383.69215	127.1987286
			Variance	43458.96777	4093.474314
Observed value	4	7
			Assumed average is poor	0
Df	3
			The t statistics	2.397079927
P(T＜＝t)one-tail	0.04806332
			t Critical one-tail	2.353363016
P(T＜＝t)two-tail	0.09612664
			t Critical two-tail	3.182449291
ID is to SC
				ID 100ng	SC 100ng
Meansigma methods	127.1987286	352.86435
			Variance	4093.474314	762.4257392
Observed value	7	4
			Assumed average is poor	0
Df	9
			The t statistics	-8.10411752
P(T＜＝t)one-tail	9.97966E-06
			t Critical one-tail	1.833113856
P(T＜＝t)two-tail	1.99593E-05
			t Critical two-tail	2.262158887
ID is to contrast
				ID 100ng	ID-PBS
Meansigma methods	127.1987286	363.97564
			Variance	4093.474314	6557.946662
Observed value	7	5

Assumed average is poor	0
			Df	7
The t statistics	-5.43722939
			P(T＜＝t)one-tail	0.000484554
t Critical one-tail	1.894577508
			P(T＜＝t)two-tail	0.000969107
t Critical two-tail	2.36462256

6.2ID and IP delivers the influence to the IL-12 anti-tumor activity

Give C57BL mouse subcutaneous (SC) inoculation 1 * 10 ⁶Individual B16F10 melanoma cells (ATCC company).Inoculate back 7 days and start treatment by intraperitoneal (IP) or intradermal (ID) administration with the IL-12 injection with two levels (10 and 100ng) in a side of carrying tumor.Repeat the IL-12 administration every other day, add up to 4 treatments.In contrast, inject the perfect dielectric (PBS) of equal volume at a side ID who carries tumor.Also move the true negative contrast with serum and tissue sample (n=15/ condition).7th, measured body weight and tumor size in 11,14,18 and 21 days.Also every one week blood sample collection be used for IL-12 and analyze.With rectangular two diameter of tumor of digital caliper measurements, and with the average tumor size to the natural law mapping of inoculation behind the tumor cell.

6.2.1ID and IP delivers the influence (10ng dosage) of IL-12 to the tumor size

Measured ID (77mm the first time after the treatment promptly the 11st day ²) growth rate and the IP (68mm of administration condition ²) the administration conditional likelihood.The 14th day, the tumor of ID treatment mice increased 56% to 120mm by previous measurement result ², and the tumor of IP treatment mice has increased 63% to 113mm ²Two treatment groups show the tumor size similar to PBS treatment group, and the latter is respectively 74mm the 11st and 14 day average tumor size ²And 126mm ²(having increased 70%).Yet the 18th day, the tumor of ID treatment mice was dwindled 6% (112mm ²), and the tumor of IP treatment mice has increased 82% (206mm ²).The 21st day, the ID condition increased 121% to 250mm ², and the IP condition has increased 27% to 262mm ²The 21st day, the average tumor size and the PBS administration animal (259mm of ID and IP condition ²) similar.Only seen in the IL-12 dosage of the 18th day 10ng that ID delivers the advantage that is better than IP.IL-12 is injected at end in the 13rd day, if treat with the relieve pain of wideer or bigger frequency, advantage may be more remarkable.

6.2.2ID and IP delivers the influence (100ng dosage) of IL-12 to the tumor size

When measure the first time after treatment the 11st day, at ID (54mm ²) in seen (70mm with respect to IP ²) the average growth rate of administration condition reduces.The 14th day, the tumor of ID treatment mice increased by 39% to 75mm by previous measurement result ², and the tumor of IP treatment mice has increased by 80% to 126mm ²Have only ID treatment group to show tumor size and the growth rate littler than PBS treatment group, the latter is respectively 74mm the 11st and 14 day average tumor size ²And 126mm ²(70%).By the 21st day, the ID condition increased by 89% to 142mm ², and the IP condition has increased by 139% to 302mm ²For the IL-12 of 100ng dosage, the average tumor growth rate of ID condition reduces with respect to IP, and the final average tumor size of ID is littler by 47% than IP.

Between ID treatment group and the IP treatment group Discrepancy Description of tumor size can the ID administration than the low dosage of IP and reach identical or better whole body and reply-save dosage and improve effect.Reduce quantity and seriousness that injection volume also should be reduced in the side effect of seeing in the higher dosage.Tumor load reduces also to mean that transfer activity reduces and survival rate raises.

6.3ID deliver the influence of position to the IL-12 anti-tumor activity

Give C57BL mouse subcutaneous (SC) inoculation 1 * 10 ⁶Individual B16F10 melanoma cells (ATCC company).Inoculate back 7 days dosage and give 100ngIL-12 injection startup treatment as ID dosage that carries tumor one side or offside.Repeat the IL-12 administration every other day, add up to 4 treatments.In contrast, inject the perfect dielectric (PBS) of equal volume at a side ID who carries tumor.Also move the true negative contrast with serum and tissue sample (n=15/ condition).7th, measured body weight and tumor size in 11,14,18 and 21 days.Also every one week blood sample collection be used for IL-12 and analyze.Gathered draining lymph node and spleen sample on the the 14th and 24 day and be used to analyze T cell, B cell and NK cell, and carry out GP100FACS and analyze.With rectangular two diameter of tumor of digital caliper measurements, and with the average tumor size to the natural law mapping of inoculation behind the tumor cell.

6.3.1 result

The 18th day, the significant difference of tumor size appearred between ID administration condition and the offside ID administration near the tumor first.75mm by the 14th day each condition ², carry out side injection (ID _Cs) tumor increased by 91% to 143mm ², and carry out tumor side injection (ID _Ts) tumor only increased by 68% to 126mm ²By the 21st day, ID _CsIncreased by 29% again, and ID _TsOnly increased by 14% to 142mm ²Two kinds of ID conditions all have and are better than the remarkable improvement that PBS handles animal, and wherein the latter is 259mm the 21st day average tumor size ²ID _TsThe final tumor size of condition compares ID _CsLittle by 23%, proved the directly advantage of the lymph relevant of targeting with tumor.

Intradermal administration 100ng IL-12 is injected at and has maximum the reduction aspect the overall tumor growth near the tumor injection position.Be not limited to concrete theory, this has proved the ability of ID injection than the effective more target tumor draining lymph node (system that relates to tumor conveying, transfer and immunne response) of other condition.The ID injection IL-12 that offside carries out is effective not as near administration tumor, but better than IP condition.Be not limited to concrete theory, this may be still not realized immunne response owing to still have arrival directly to relate to tumor lymphatic by direct targeting lymph.

6.4 use ID to deliver the targeting lung tissue

Tissue and cyclical level by measure R SV specific monoclonal antibody 48 (RSV MAB 48) have compared intramuscular (IM) and ID delivery approach.RSV MAB 48 generates by the RSV fusion protein immunization Balb/c mice with purification.Gather (primed) B cell of sensitization, and merge by PEG method and 653 myeloma system.Those of ordinary skills can use technology well-known in the art to obtain other suitable monoclonal antibody.

Gather lung tissue and measure the situation that exists of antibody.Gathered the lung sample in back 3 hours, 24 hours, 7,14,21 and 28 days in injection.In order to finish predetermined sampling, research divides six stages to carry out.Each tissue sampling time and road specify two to repeat or two animals, thereby provide two duplicate samples for the sampling of each time point.

6.4.1 detailed process

6.4.1.1 preparation is used for the lung tissue lysate of ELISA

The results lung, rinsing in 0.9%NaCl, in-20 ℃ freezing, and at Vacutainer ^Be stored in-20 ℃ (5ml) until check.Before the cracking, will organize thawing, and, remove any residual blood by the lung tissue surface with HanksShi balanced salt solution (4ml) and the thorough rinsing of pipettor.Then by sucking-off Hank sShi balanced salt solution in the pipe and abandon.In every batch of lung, add 1mlCHO 2x lysis buffer (contain 1mM Phenylmethanesulfonyl fluoride, 0.25M Tris (methylol) aminomethane hydrochloric acid pH with pH/mV/ thermometer and sodium hydrate particle transfer to 8.0,0.05M sodium chloride, 0.5%Nonidet ^P40 replacement liquid, 0.5% NaTDC).With Vacutainer ^Keep stable, fall speed change homogenizer and enter lung solution, make blade be positioned at lung piece top.Speed change homogenizer is started about 20-30 second, destroy lung tissue fully.Carefully avoid excessive homogenate or produce the heat of the antibody of may degrading.Lung tissue solution after the homogenate is transferred to Falcon ^BlueMax ^TMBe used for supersound process in the 15ml polystyrene plastics conical pipe.Material is about 30 seconds supersound process on ice 3 times.Then homogenate, lung tissue suspension after ultrasonic are transferred in the miniature centrifuge tube, and in Marathon Micro H Fisher Scientific micro centrifuge centrifugal 15 minutes.Then, shift out supernatant and be stored in-20 ℃ until protein and antibody test.Abandon precipitation.

6.4.1.2 bioassay standard

The mensuration of gross protein is to use BCA (bicinchoninic acid) reagent (Pierce, catalogue #23225) to carry out.

The bioassay standard thing is following generation: do not gather lung tissue by treating cotton mouse (not accepting antibody), cracking as mentioned above, totally be divided into 5 pipes with what reclaim, and mix the RSV MAB 48 of dose known amounts (being respectively 300 μ g/ml, 30 μ g/ml, 3 μ g/ml and 300ng/ml) to 4 pipes.Remaining that pipe does not add MAB as negative control.Each spiked sample and null value control material are diluted with the ELISA sample loading buffer, produce by 1: 10 ²To 1: 10 ⁵The log10 dilution thing.

6.4.1.3ELISA measure

The anti-mice IgG2a of Zymed rabbit liquid storage (500 μ g/ml, catalog number 61-0200, lot number 20168583) is cushioned liquid (Sigma) with the carbonate bag is diluted to 3 μ g/ml., covered and by the hole of 96 orifice plates (just inner 60) with the anti-mice IgG2a of 100 μ l3 μ g/ml Zymed rabbits carbonate solution bag at CO ₂Left standstill in the incubator 1 hour.By abandoning coating buffer in the hole, and plate bounced on napkin to remove remaining coating buffer.The PBS/ polysorbas20 (Sigma P3563) that contains 5% milk powder with 250 μ l seals non-specific site.Use the Parafilm overlay, and at CO ₂2 hours (37 ℃) of insulation in the incubator.Abandon confining liquid, and plate is cleaned 3 times with the PBS/ polysorbas20, on napkin, bounce then to remove remaining cleanout fluid.Sample and RSV 48.4.1 contrast (is anti-) are suitably diluted with the conformance with standard curve.Standard curve is by the RSV MAB 48 and blank composition the in 300ng/100 μ l hole, 100ng/100 μ l hole, 30ng/100 μ l hole, 10ng/100 μ l hole, 3ng/100 μ l hole, 1ng/100 μ l hole, 0.3ng/100 μ l hole.Sample is diluted in containing the PBS/ polysorbas20 of 0.5% milk powder, and 2 of each dilution factors repeat and each hole 100 μ l.Then with plate at CO ₂Insulation is 1 hour in the incubator.

Abandon an anti-solution, plate is cleaned 3 times with the PBS/ polysorbas20, and on napkin, bounce to remove remaining cleanout fluid.In each hole, add 100 μ l HRP conjugates (set of goat anti-mouse conjugate) then.In order to prepare HRP conjugate set, in containing the PBS/ polysorbas20 solution of 0.5%w/v milk powder, 30ml adds anti-IgG2a of 2 μ l (Southern Biotech) and the anti-IgG of 4 μ l (Sigma).In each hole, add 100 μ l solution.With the plate that covers at CO ₂1 hour (37 ℃) of insulation in the incubator.Abandon two anti-solution then, plate is cleaned 4 times with the PBS/ polysorbas20, and on napkin, bounce to remove remaining cleanout fluid.In each hole, add 100 μ l TMB (SigmaT8665) substrates, and developed the color in the dark 30 minutes.In each hole, add 200 μ l 0.5MH ₂SO ₄, and read the absorbance of 450nm.

By primary OD value operation (among the Excel of Microsoft) forecast function is measured the actual weight of antibody.The forecast function is by existing numerical computations or predict following numerical value.For example, be the y value to the predictive value of specifying the x value.Known numerical value is existing x value and y value, by the new numerical value of linear regression prediction.

6.4.1.4 model antibody

This single channel ID is mouse IgG 2a to employed antibody (RSV MAB 48) in the IM research.RSV MAB 48 identification fusion rotein, and in the tissue culture experiment, show the blocking-up infection.This antibody of amplification in ascites, with Zymed protein A column purification, dialysis is removed excipient and is not damaged the dissolubility of antibody with as much as possible in 1/10PBS.

6.4.1.5 dosage

Every animal is accepted injection with the 15mg/kg body weight.Consider that excipient almost accounts for the weight of half, the antibody dosage of the actual acceptance of cotton mouse is about 7.5mg/kg.

The calculating of the sample of IM and ID injection shows below:

Cotton mouse body weight=98.9 grams;

In order to use 75 μ l antibody-solutions with 15mg/kg, need the concentration of 1.48mg/75 μ l;

In order to comprise the injection loss, get 3.94mg and be dissolved in 200 μ l, and administration 75 μ l.

6.4.1.6 delivery apparatus and method

Use 30G BD pin to carry out IM and ID injection.IM injects following carrying out: clamp (pinching) back leg muscle, produce the degree of depth; Make the inclined-plane can imbed muscle the pin certain angle that tilts, deliver the IM volume injected, and in muscle tangibly.ID injects following carrying out: enter with shallow as far as possible angle, the upset inclined-plane upwards produces in " blister " before injection then.

6.4.2 result

According to back 3 hours, 24 hours, 7,14,21 and 28 days measurement results of injection, when checking for the first time, inject back 3 hours (Fig. 4) and in the lung tissue that obtains by ID dispenser animal, observed of the rising of organ antibody with respect to IM dispenser animal to lung tissue.The time point of observing this rising for the last time is back 21 days of injection.Injected back 28 days, and between ID and IM dispenser animal, do not observe the difference of organ antibody horizontal.

Injection back 3 hour records are to the maximum difference percentage rate, and ID dispensing this moment animal shows that the antibody in their lungs is higher by 350% than IM dispensing animal.Adding up to the overall rising of all measurement results of six time points is about 18% (about 57ng of ID dispenser animal is to about 48ng of IM dispenser animal).In addition, in 3 hours ID samples indication lung antibody to begin effect faster, and the higher peak level (C of antibody in 1 all ID sample indication lungs _Max) higher.

These digital proofs ID delivers can reach higher antibody horizontal in target tissue (lung), and need not to improve the administration quantity of antibody.Therefore, compare with the intramuscular approach, the needed antibody of protection of intradermal delivery realization similar level still less.Except cost advantage, reduce if in lung, keep target trough value desired material, the risk that causes adverse side effect (such as human anti-mouse antibody (HAMA)) so reduces.Be not limited to concrete theory, the raising of bioavailability may be because injection institute at tissue site promotion reactive compound better absorption and/or reduce to the unfavorable degraded of medicine minimum.

6.5 folding divides (split) dosage two approach researchs

6.5.1 experimental design

The preliminary bioavailability study in the inside of carrying out in cotton mouse (in-house) has proved that direct intranasal administration antibody can produce higher peak level in lung tissue.This observed result has caused splitting the formation of the two approach strategies of dosage, and it is used for not only obtaining the therapeutic antibodies level but also obtain preventative antibody horizontal.This research is carried out in Balb/c mice (group n=5).

At first carried out a series of preliminary experiments and delivered volume and sampling time subsequently to determine best IN.As described in 6.4 joints, carry out the collection of lung material, just asynchronism(-nization).Since the animals received in this research the different special monoclonal antibodies of RSV (Palivizumab), so the ELISA of pulmonary docimasia and 6.4 to save described method different.In these preliminary experiments, each is organized the Balb/c mice and divides three IN volume of taking medicine to accept Synagis Palivizumab with the 15mg/kg body weight.1,3 and 5 hour collection lung tissue after the administration.Each test and matched group use 5 mices.All lung tissues that collection is gathered by every animal, homogenate in lysis buffer by centrifugal clarification, and is added to supernatant in the elisa plate, has wherein fixed the RSV-FP peptide.The lung homogenate thing that merges in test or the matched group is used for measuring.

6.5.2ELISA flow process

Exemplary ELISA flow process below using is carried out ELISA:

1. by taking out 1mg/ml RSV F 64 polymers peptide stock solutions in the refrigerator.Liquid storage is cushioned liquid (Sigma) with the carbonate bag is diluted to 10 μ g/ml;

With 100 μ l, 10 μ g/ml RSV FP, 64 polymers bags by the hole of 96 orifice plates (just inner 60).Cover and at CO ₂Left standstill in the incubator 1 hour;

With coating buffer by abandoning to tank in the hole, and plate bounced on napkin to remove remaining coating buffer.The PBS/ polysorbas20 (Sigma P3563) that contains 5% milk powder with 250 μ l seals non-specific site.Use the Parafilm overlay, and at CO ₂2 hours (37 ℃) of insulation in the incubator;

4. confining liquid is discarded in the tank, and plate is cleaned 3 times with the PBS/ polysorbas20.Plate is bounced on napkin to remove remaining cleanout fluid;

5. dilute sample is with the conformance with standard curve.For the drawing standard curve, use the Palivizumab in 300ng/100 μ l (hole), 100ng/ hole, 30ng/ hole, 10ng/ hole, 3ng/ hole, 1ng/ hole, 0.3ng/ hole, and blank.Sample is diluted in containing the PBS/ polysorbas20 of 0.5% milk powder.Each dilution factor carries out 2 repetitions.In each hole, add 100 μ l.With plate at CO ₂Insulation is 1 hour in the incubator;

6. an anti-solution is discarded in the tank, and plate is cleaned 3 times with the PBS/ polysorbas20.Plate is bounced on napkin to remove remaining cleanout fluid;

7. in each hole, add 100 μ l HRP conjugates (the anti-people Ig of goat).In order to prepare HRP conjugate work liquid storage, in containing the PBS/ polysorbas20 solution of 0.5%w/v milk powder, 30ml adds the anti-people Ig of 2 μ l (Promega).The vortex concussion also adds 100 μ l in each hole.With the plate that covers at CO ₂1 hour (37 ℃) of insulation in the incubator;

8. two anti-solution are discarded in the tank, and plate is cleaned 4 times with the PBS/ polysorbas20.Plate is bounced on napkin to remove remaining cleanout fluid;

9. in each hole, add 100 μ l TMB (Sigma T8665) substrates, and developed the color in the dark 30 minutes; And

10. in each hole, add 200 μ l 0.5M H ₂SO ₄, and read the absorbance of 450nm.

By original OD value operation (among the Excel of Microsoft) forecast function is measured the actual weight of antibody.

6.5.3 delivery apparatus and method

The IM injection reuses 30G BD pin (re-ordering #305106) and carries out.Prepare the IM injection site by clamping the back leg muscle to produce the degree of depth.Make the inclined-plane can suitably imbed muscle the pin certain angle that tilts.Deliver volume injected and tangibly in muscle.

The IN administrable has been installed the P200 Ranin pipettor of disposable tip and has been used.

6.5.4 result

Fig. 5 has shown and the take medicine stable increase of the consistent lung antibody concentration of the increase of volume of intranasal.The antibody that IN delivers is imitated.Yet the level than any other delivery approach record is high 2 times at least for detected antibody peak concentration in 100 μ l group.These are found with consistent from the observed result in the early time of cotton mouse research.Although the antibody that IN delivers is removed rapidly, yet at antibody concentration and organ volume that this institute relates to, only need there be the time of a few minutes in the antibody with normal affinity.

Utilize above-mentioned discovery, the bioavailability of the Palivizumab that IM is delivered and the dosage that splits between IN and IM approach compare.Balb/c (n=5) mice is accepted Synagis Palivizumab with the 15mg/kg body weight.All IM and IN volume injected all are 75 μ l.Gather lung tissue in 1 hour after the administration, and after 1 week, gather lung tissue once more.All lung tissues that collection is gathered by every animal, homogenate in lysis buffer, and by centrifugal clarification, as above 6.4 joints are described.All clarifying homogenates are carried out BCA protein check, thus can be on the protein basis that equates compare test and control tissue.Merging is from the clarification lung tissue homogenate of each group, and is added in the hole of the elisa plate of RSV-FP peptide bag quilt.Fig. 6 provides the example of expectation and non-expected result.

Fig. 7 has shown that splitting dosage 15%IN/85%IM produces the lung antibody of substantial level immediately, and during 1 week the essential preventative level that exists without any reduction.The reliability index of lasting level when the competitive level of antibody is 1 month when before determining for 1 week.The antibody amount of finding in the lung tissue of being gathered by the IN/IM group is consistent with the antibody concentration that neutralization in external check is infected.On the contrary, but standard I M administration did not produce the antibody of detection limit in the time of 1 hour.The lung antibody horizontal of IM and IN/IM group is suitable when 1 week.Fig. 8 has shown that the antibody concentration of 6 μ g/ml can be viral in a large number at extracorporeal blocking.

6.5.4.1 after the administration one hour

As shown in Figure 4, split the two approach of dosage and deliver the tactful treatment concentration of Synagis Palivizimab in lung tissue that causes greater than the homogenate of 6 μ g/ml clarification lung tissue.On the contrary, standard I M delivery Synagis Palivizimab does not cause any detectable antibody in the lung homogenate.

A 6.5.4.2 week after the administration

The fractionation approach is delivered strategy and is caused the concentration of Synagis Palivizimab in lung tissue greater than 100ng/ml clarification lung tissue homogenate (standard I M delivers the preventative antibody horizontal that reaches usually).

Two approach strategies have reached the treatment level of antibody in lung tissue, and do not meet with the loss of longer-term prevention level.This all is to realize with the FDA approval dosage of standard.Can be in the identical time, give sick special antiviral agents of RSV and antibiotic medicine in the two approach modes of identical fractionation dosage.

6.6 virus attack

In the further research, the inventor has proved the existing infection of can disappearing of standard P alivizumab dosage that 15% IN delivers.With 1.3 * 10 ⁵Individual plaque forming unit is attacked mice.Infected back three hours, the test group is carried out the IN treatment with the Palivizumab (about 2.25mg/Kg) of single dosage, and matched group is accepted saline.Infected back three days, and gathered lung tissue and homogenate.The homogenate that obtains is assigned on the Hep-2 monolayer, and to the viral sign of culture monitoring survival.Reclaiming the total homogenate volume that obtains by every animal is about 1.5-2ml, is 100 μ l and place the amount of each instrument connection.Following table has shown the Synagis IN infection of having disappeared, and does not observe plaque forming unit.

Handle	Infect the lung tissue (directly measuring) of gathering in back 72 hours
		With IN saline treatment animal	The 29PFU/ hole
Handle animal with IN Synagis (15% dosage)	Do not observe PFU

Claims (33)

1. be used for having needed human experimenter to treat method for cancer, comprise the intradermal zone that at least a therapeutic agent is delivered to human experimenter's skin, wherein with deliver this therapeutic agent by the approach beyond the intradermal delivery and compare, this therapeutic agent causes the bigger reduction of tumor growth.

2. be used for treating method for cancer the human experimenter, comprise the intradermal zone that at least a therapeutic agent is delivered to human experimenter's skin, wherein with deliver this therapeutic agent by other approach beyond the intradermal delivery and compare, this therapeutic agent causes the prolongation of human experimenter's life-span intermediate value.

3. claim 1 or 2 method, the approach beyond the wherein said intradermal delivery is subcutaneous delivery.

4. claim 1 or 2 method, the approach beyond the wherein said intradermal delivery is that intramuscular is delivered.

5. claim 1 or 2 method, the approach beyond the wherein said intradermal delivery is that intravenous is delivered.

6. claim 1 or 2 method, the approach beyond the wherein said intradermal delivery is that epidermis is delivered.

7. claim 1 or 2 method, wherein cancer is selected from down group: lymphoma, leukemia, breast carcinoma, melanoma, pulmonary carcinoma, renal carcinoma and colorectal carcinoma.

8. be used for method at least a therapeutic agent of human experimenter's administration, comprise the intradermal zone that therapeutic agent is delivered to human experimenter's skin, make and deliver this therapeutic agent by other approach beyond the intradermal delivery and compare that this therapeutic agent has higher tissue biological's availability in particular organization.

9. the method for claim 8, wherein the drug treatment agent is the disease that is selected from down group in order to treat: cancer, cancerometastasis, tumor growth or infectious disease.

10. for the method for prevent disease at least a therapeutic agent of human experimenter's administration, comprise the intradermal zone that this therapeutic agent is delivered to human experimenter's skin, make and deliver this therapeutic agent by other approach beyond the intradermal delivery and compare that this therapeutic agent has higher tissue biological's availability in particular organization.

11. the method for claim 10, wherein Yu Fang disease is selected from down group: cancer, cancerometastasis, tumor growth or infectious disease.

12. for the outbreak that postpones morbid state or progress method at least a therapeutic agent of human experimenter's administration, comprise the intradermal zone that therapeutic agent is delivered to human experimenter's skin, make and deliver this therapeutic agent by other approach beyond the intradermal delivery and compare that this therapeutic agent has higher tissue biological's availability in particular organization.

13. the method for claim 12, wherein morbid state is selected from down group: cancer, cancerometastasis, tumor growth or infectious disease.

14. claim 1,2,8,10 or 12 method, wherein therapeutic agent is selected from down group: anticarcinogen, antitumor agent and chemotherapeutant.

15. claim 1,2,8,10 or 12 method, wherein therapeutic agent is selected from down group: antibody, vaccine and cell therapy.

16. the method for claim 15, wherein antibody is selected from down group: preventative antibody, polyclonal antibody, monoclonal antibody, murine antibody, people's antibody, chimeric antibody or antibody fragment.

17. claim 1,2,8,10 or 12 method, wherein therapeutic agent is selected from down group: angiogenesis inhibitor, cytokine or chemotactic factor.

18. the method for claim 17, wherein cytokine is interleukin or interferon.

19. the method for claim 18, wherein interleukin is an il-1 2.

20. claim 1,2,8,10 or 12 method are wherein by pin or sleeve pipe delivery of therapeutics agents.

21. claim 1,2,8,10 or 12 method wherein are inserted into pin or telescopic outlet the degree of depth of the about 3mm of about 300um-.

22. claim 1,2,8,10 or 12 method, wherein pin or telescopic specification are 30-36 number.

23. claim 1,2,8,10 or 12 method, wherein pin or telescopic specification are 31-34 number.

24. claim 1,2,8,10 or 12 method, wherein use at least one small size hollow needle delivery of therapeutics agents of outlet exposure height 0-1mm, the degree of depth of skin to 0.3mm-2mm inserted in described outlet, made material deliver the degree of depth that occurs in 0.3mm-2mm.

25. claim 8,10 or 12 method, wherein particular organization is selected from down group: lymphoid tissue, mucosal tissue, lymph node, skin histology, germinal tissue, cervical tissue, vagina tissue, lung, spleen, colon, thymus, bone marrow, hemolymph sample tissue and lymphoid tissue.

26. the method for claim 25, wherein lymphoid tissue is selected from the relevant lymphoid tissue of epithelium, mucosa-associated lymphoid tissue, elementary lymphoid tissue, secondary lymphoid tissue.

27. claim 8,10 or 12 method have accumulated the about 30ng therapeutic agent of about 10pg-in wherein per 50 μ g particular organizations.

28. claim 8,10 or 12 method have accumulated the about 15 μ g therapeutic agents of about 10pg-in wherein per 50 μ g particular organizations.

29. claim 8,10 or 12 method have accumulated the about 30ng therapeutic agent of about 1cg-in wherein per 50 μ g particular organizations.

30. be used for having needed human experimenter to treat method for cancer, comprise the intradermal zone that at least a therapeutic agent is delivered to human experimenter's skin with previously selected dosage, wherein compare with the dosage of delivering by other approach beyond the intradermal delivery, this previously selected dosage has reduced at least half.

31. the method for claim 30, wherein this therapeutic agent is an il-1 2.

32. be used for having needed human experimenter to treat method for cancer, comprise the intradermal zone that at least a therapeutic agent is delivered to human experimenter's skin, make and deliver identical treatment agent by other approach beyond the intradermal delivery and compare that it is faster that this therapeutic agent begins effect.

33. the method for claim 32, therapeutic agent wherein are il-1s 2.

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