JP2003524646A - Pharmaceutical aerosol formulation - Google Patents

Abstract

  (57) [Summary] A pharmaceutical formulation is disclosed. The formulation comprises: a therapeutically effective amount of a protein or peptide drug, a fluid for containing the drug having a molecular size of 1 kilodalton to about 150 kilodaltons, ie, a fluid carrier for containing the drug, and an amino acid; It contains a stabilizer selected from its derivatives, or mixtures thereof.

Description

Detailed Description of the Invention

This application claims priority based on US Provisional Patent Application No. 60 / 177,987, filed January 25, 2000.

[0002] FIELD OF THE INVENTION The present invention relates to pharmaceutical aerosol formulations and, more particularly, relates to a pharmaceutical aerosol formulation comprising a protective colloid stabilizer.

Related Art Delivery of drugs to the lungs by inhalation is a common local condition such as cystic fibrosis, bronchial asthma, and chronic obstructive pulmonary disease, as well as pain management, immunodeficiency, hormone therapy, It is an important means for treating a variety of conditions, including some systemic conditions including erythropoiesis, diabetes and the like. Steroids, β2 agonists, anticholinergic agents, proteins, and polypeptides are among the agents administered to the lung for such purposes. Such agents are usually administered to the lung in the form of an aerosol consisting of particles of a size suitable for inhalation (diameter less than about 10 μm). To ensure the proper particle size in the aerosol, the particles are prepared in a size suitable for inhalation, and then pressurized gas in the case of a pressurized metered dose inhaler (MDI), or a dry powder inhaler (DP
It can be added in the colloidal dispersion containing any of the air in case I).
Alternatively, the formulation may be prepared in the form of a solution or emulsion to avoid concerns about the proper particle size in the formulation. However, solution formulations must be dispensed to create particles or droplets of suitable size for inhalation.

For MDI preparations, the aerosol formulation once prepared is packaged in an aerosol closed container with metered dose valve. In the patient's palm, the formulation is dispensed via a drive adapted to direct the dose from the valve to the patient.

It is important that the aerosol formulation is stable so that the delivered dose released from the metered dose valve is reproducible. Rapid creaming, settling, or flocculation after agitation is a common cause of non-reproducible quantity in suspension formulations. When using a two-mixed aerosol formulation containing only a drug and a propellant such as 1,1,1,2-tetrafluoroethane, or when using such a formulation further containing a trace amount of a surfactant. , Especially so. The sticking of the valve may also cause the inability to reproduce the quantity. To overcome such problems, MDI aerosol formulations often include a surfactant, which acts as a suspending agent to stabilize the suspension for a sufficient time to provide a reproducible distribution. Some surfactants also function as lubricants that smooth the valve to ensure smooth operation. A myriad of substances are known and disclosed for use as dispersants in aerosol formulations. However, the suitability of a substance depends on the particular drug and propellant or type of propellant used in the formulation.

[0006] It can be difficult to dissolve sufficient amounts of conventional surfactants in hydrofluorocarbon (HFC) propellants such as HFC-134a and HFC-227. To overcome that problem, cosolvents such as ethanol have been used, as described in US Pat. No. 5,225,183. Another attempt to avoid the use of cosolvents is to use substances such as those which are either readily soluble or evenly dispersible in hydroxyfluorocarbon propellants and which are said to be effective surfactants or dispersants in aerosol formulations. Including. Such materials include certain fluorinated surfactants and certain polyethoxy surfactants.

[0007] Relatively small molecules are much more predictable in terms of aerosol formulation properties than macromolecules. Macromolecules such as peptides or proteins with molecular sizes of, for example, about 1 kilodalton to about 150 kilodaltons are very unpredictable and are unique in forming stable aerosol formulations and providing reproducible doses. Indicates the problem.

Most peptide and protein drugs, such as hormones such as insulin, amylin, enzymes, anti-infectives, are highly variable in their amino acid composition and three-dimensional structure. Consequently, its surface activity is highly variable and, importantly, is based on most of its basic and structural properties such as molecular weight, adsorption power, solubility, partition coefficient, and isoelectric point. No model is available yet to explain the differences in protein surface activity. For example, hemoglobin has a higher affinity for solid surfaces than albumin, but the molecular weights of the two proteins are very similar. Basically, the difference in surface activity of peptides and proteins comes from the primary sequence of amino acids that uniquely characterizes each type of protein. Amino acid side chains often change dramatically, in that some amino acid side chains are uncharged at any pH but exhibit significant polar character (serine, threonine). Other amino acids are ionic, and from fairly acidic functional groups (aspartic acid, glutamic acid are completely negatively charged at physiological pH (7.4)), for example histidine (
changes to basic functional groups such as imidazole groups (partially positively charged at pH 7.4) and even more basic amino groups of lysine and arginine which are fully positively charged at pH 7.4 . Another group of amino acids with somewhat hydrocarbon-like characteristics (tryptophan, phenylalanine, isoleucine, etc.) appears to be generally less soluble in water than many other amino acids found in biological systems. It is worth noting that the hydrophobicity of these water-phobic amino acids is significantly altered by their specific structure in proteins. For example, a single methyl group side chain of alanine is only 0.5 kcal / min for the free energy to transfer from water to the organic phase.
However, the bicyclic indole group of tryptophan is 3.4k.
contribute to cal. The differences in amino acid side chains, along with the many different types of chemical interactions that occur in solution and at the surface, significantly contribute to the stability of aerosol formulations and their transport of peptide and protein biotherapeutic agents across biological membranes. Is expected to have the effect of

Various features of amino acid side chains, along with the complexity of the combination of various amino acids in each particular protein, physicochemical properties of proteins, their intermolecular and intramolecular reactivity, and surface interactions. This means that your ability to do so will change considerably.
Due to its large size, as well as due to the large number of charged amino acid side chains, proteins have many charges distributed over their outer surface. This would result in enormous changes in the stability of aerosol formulations and their pulmonary uptake. Peptide and protein drugs usually have multiple ionization sites and thus their formulations show a pH-dependent dissolution profile. Importantly, the hydrophilic nature of these compounds provides excellent conditions for high water solubility. As a result, most peptide and protein drugs exhibit fairly poor lipid solubility properties, which is probably that their dispersion in hydrofluorocarbon propellants is physically and chemically stable over a wide range of conditions. That might be one reason. Aerosol formulations include peptide and protein drugs in a carrier or formulation medium, and the substantial insolubility of the drugs in the medium results in hydrolytic and chemical deactivation, which is usually common in aqueous solutions. It is necessary to reduce.

The large surface area, absorbable carriers, and extensive vasculature constitute a favorable absorption environment for proteins and peptides when transported by the pulmonary route. Studies have shown that intratracheal (it) administration of peptides is rapid and quantifiable; the resulting distributions are often localized to the central airways. Administration by aerosol, which depends on, for example, particle size distribution, can be used to result in a more uniform distribution with more alveolar entry. Drug absorption from the respiratory tract is based on the site of deposition, the method of drug transport, the type of solute presentation, and the composition of the formulation. Thus, formulation and device characteristics will dramatically affect the rate and extent of peptide absorption from the lungs. Studies have shown that the absorption rate of low molecular weight compounds after aerosol transport is almost double that of it transport. Providing peptides and proteins as a hydrophobic dispersion via a multi-dose inhaler (pMDI), resulting in more entry of drug particles into the peripheral lungs, using the aqueous drip method in the lungs It is desirable that the drug is remarkably absorbed more than when the drug is centrally deposited.

The fact that insulin can be absorbed from the lungs into the bloodstream has been proven by many scientists. A 1990 review paper [Lung, supplement pp. 677-684] shows from multiple studies that aerosolized insulin delivered to the lung has a half-life of 15-25 minutes, but results vary considerably. It was A comprehensive study showed that bioavailability of 5.6% for liquid insulin infused into the central respiratory tract compared to more than 50.7% for aerosolized insulin administered peripherally to the lungs of rabbits. It has been shown to bring. Therefore, those studies require peripheral delivery of aerosolized insulin to the lungs to yield maximum efficiency,
In addition, we support the argument that inadvertent central deposition of inhaled aerosolized insulin would result in a 10-fold lower effect than desired. Such a 10-fold change in dosing is clearly unacceptable if aerosolized insulin should be an effective means of treating diabetes. Therefore, there is a need for an effective and very precise aerosol device to achieve the acceptance required to administer insulin to human patients. The above concepts for using aerosolized insulin in the management of diabetes also apply to the insulin partner hormones amylin and glucagon in the regulation of plasma glucose concentrations, which until now had to be administered by subcutaneous injection (sc). It

[0012] Dry powder delivery of peptide and protein drugs has unique advantages in formulation that do not occur with liquid delivery such as pMDIs and nebulized solutions. Due to the improved solid state stability, dry powder aerosols of peptide and protein drugs are attractive from a formulation point of view since many of the undesired interaction effects of solution and liquid states are avoided. In this regard, Ru
See US Pat. No. 5,672,581 to bsamen et al. and US Pat. No. 5,775,320 to Patton et al.

Although both Rubsamen and Patton's attempts are therapeutically feasible, their complexity and the estimated inherent cost limit their application to the treatment of chronic diseases such as diabetes. Therefore, it is problematic to use expensive and complex devices such as portable electronic nebulizers to routinely bring hypoglycemia to patients in need.
Further, it is a problem to use dry powder aerosol devices, such as the Patton device, to transport hypoglycemic drugs to the body through the lungs, which are large, bulky and difficult to clean. Therefore, the first purpose in formulating peptide or protein drugs as dry powder inhalation aerosols (DPIs) is to
An aerocolloid (aer) in which the added excipients are chemically and physically stable and can remain suspended until the drug particles reach the alveoli or other absorption sites.
ocolloid). Once at the site of absorption, drug particles are efficiently trapped at the site of deposition, rapidly dissolved in epithelial lining fluids and further absorbed across biological membranes, and thus by metabolic enzymes in the respiratory tract. Potential deactivation is limited.

Spray drying is a process used to prepare drug particles for drug formulation. Spray drying constitutes a single step that transforms a solution or suspension into a uniform powder. Normally, spray drying forms spherical particles, which are often hollow, thus resulting in a powder having a low bulk density compared to the starting material. The powder properties of the spray dried material (ie particle size distribution, bulk density, porosity, water content, dispersity, etc.) are usually good in many respects, but the particles produced by the method have poor flow properties. Show the characteristics. Moreover, spray drying is less desirable for heat sensitive compounds such as peptide and protein drugs, as the process requires heating during particle formation. Therefore, it is problematic that most dry powder aerosols exhibit sticking and poor flow through the device hardware to the extent that dose delivery accuracy is problematic to the patient.

Another problem associated with peptide and protein formulations as dry powder aerosols is to device the material as a mass so that the mass breaks during aerosolization, releasing individual particles before entering the respiratory tract. It is to pack in. The preparation of strong agglomerates of micron or submicron sized particles is a fairly straightforward task that can be accomplished with conventional granulation techniques, with or without polymeric binders.
However, the need for the clumps to break up into the first particles upon entry into the respiratory tract is probably due to interparticle forces being too great to allow easy and efficient de-agglomeration, and thus premature granulation. Would hinder a simple conventional approach for. The total adhesive force between two different particles or the total cohesive force between two similar particles can be considered to consist of the sum of one or more attractive forces. Many of these forces are known to be responsible for the formation of adhesive units between the dry powder and excipient particles in the formulation. Thus, the purpose of any manipulation of interparticle forces is strong enough to withstand flow, storage, and packing in transport devices, but can be rapidly and completely deaggregated by shear stress in the flow of inspiration. It will form lumps of between about 50 and 200 μm. The problem, which is quite common in peptide and protein aerosol formulations, can be completely avoided in liquid formulations in which the drug is insoluble, is provided as a colloidal dispersion and is sterically protected against self-binding. Therefore, it is desirable to formulate peptide and protein drugs as loosely aggregated colloids in non-aqueous media such as hydrofluorocarbons, which when aerosolized into the respiratory tract rapidly and easily break into individual particles. Furthermore, it is desirable to provide peptide and protein drugs in a formulation system in which the drug particles have a permanent random movement, thus eliminating the aggregation of individual drug particles.

Other non-injectable anti-diabetic drugs have been proposed, some of which, when formulated with surfactants and other membrane-penetrating agents, produce nasal insulin followed by a biotherapeutic response. Have been shown to bring (Moses et al., Diabetes, Vol. 32,
November 1983; Salzman et al., New England Journal of Medicine, Vol. 312
, No.17). Significant variability between subjects and varying degrees of nasal membrane irritation were observed. Diabetes mellitus is a chronic disease that needs to be treated continuously by insulin administration, and further, it is nasally administered non-invasively because it tends to increase mucosal irritation after repeated exposure to a membrane penetration enhancer. Attempts to develop insulin have not been commercialized. Therefore, a safe, reproducible and efficient non-invasive vehicle for pulmonary peptide and protein drugs such as MDIs is desirable and needed.

[0017] Surprisingly Summary of the invention, for example, without any use of surfactants, such as the co-solvent or low molecular sorbitan trioleate is added to the two mixtures aerosol formulations of drugs, such as ethanol, It has been discovered that new and stable pharmaceutical aerosol formulations of polymeric drugs can be obtained. The use of a protective colloid stabilizer results in a stable pharmaceutical aerosol formulation.

[0018] DETAILED DESCRIPTION OF THE INVENTION This application, filed Sep. 22, 1998 U.S. Patent Application Serial No. 09 / 158,369 (by reference all of which are incorporated herein) refers to.

The present invention relates to a stable suspension aerosol formulation suitable for pressurized delivery comprising (1) a particulate polymeric drug or drug, (2) a suitable propellant, and (3) a suitable stabilizer. .

Suitable polymeric agents are those suitable for administration by inhalation, used in oral and nasal inhalation therapy. For example, air, hydrocarbon gas, chlorofluorocarbon (C
FC) stable colloidal dispersions of drugs in fluids such as propellants, or non-CFC propellants such as tetrafluoroethane (HPA-134a) and heptafluoropropane (HFA-227) are described.

Stabilizers of multiionic species such as amino acids and low molecular weight peptides are used as non-active pharmaceutical ingredients which trigger the loss of adhesive bonding between drug particles. Thus, electret or sterically stabilized aerocolloid particles of a given drug are formed. The electret is an electrostatic equivalent of a permanent magnet.
quivalent), but fragile in the presence of moisture, such as is present in air or in ambient humidity conditions of the respiratory tract. Accordingly, the present invention applies to dry powder aerosols, portable nebulizer systems, as well as pressurized metered dose inhalers.

The resulting aerocolloid is chemically and physically stable and is suspended until a given drug particle reaches the alveoli or other absorption site in the respiratory tract of the patient, eg, the human or animal being treated. It can continue to be cloudy. Once at the absorption site,
Ambient water efficiently traps drug particles at the site of deposition, rapidly dissolves in intraepithelial fluid, and is rapidly absorbed across the patient's biological membrane, thus potentially deactivating by metabolic enzymes in the respiratory tract. Is limited.

Agents suitable for the present invention are agents that form a stable hydrophobic dispersion suitable for delivery to a patient such as a human or animal. Usually, such drug is 0.5
Included are peptide, polypeptide, or protein biopharmaceuticals having a molecular weight of kilodaltons to 150 kilodaltons. Specifically, as a peptide, polypeptide, or protein biopharmaceutical, a diabetic aid; insulin and insulin analogs; amylin; glucagon; surfactants;
Interleukins such as interleukin-1, interleukin-2, and cytokines such as interferon, immunomodulatory peptides such as chemokines, lymphokines; taxol; erythropoietin; thrombolytics and heparin; antiproteases, antitrypsin, and amyloid; rhDNase; antibiotics and other anti-infectives; hormones and growth factors such as parathyroid hormone, LH-RH, and GnRH analogs; nucleic acids; DDAVP; calcitonin; cyclosporine; ribavirin;
Enzymes; heparin; hematopoietic factors; cyclosporine; vaccines; immunoglobulins; vasoactive peptides; antisense drugs; genes, oligonucleotides, nucleotide analogs and the like.

The term diabetic aids includes natural, synthetic, anti-synthetic and recombinant agents such as activin, glucagon, insulin, somatostatin, proinsulin, amylin and the like.

The term “insulin” is meant to include natural extracted human insulin, recombinant human insulin, insulin extracted from bovine and / or porcine, recombinant porcine and bovine insulin, and mixtures of any of these insulin products. Be interpreted. The term typically includes polypeptides that are used in the treatment of diabetes in a substantially purified form, but is also intended to include use in the form of a commercially available pharmaceutical with additional excipients. There is. Insulin is preferably made recombinantly and can be dried (completely dried) or in solution.

The terms “insulin analog”, “insulin monomer” are used interchangeably herein, in which one or more amino acids in a polypeptide chain have been replaced by another amino acid, And / or any form of "insulin" as defined above, wherein one or more amino acids have been deleted, or one or more additional amino acids have been added to the polypeptide chain or amino acid sequence, It is intended to include those that act as insulin in lowering blood glucose levels. Generally, "insulin analogs" of the invention refer to "insulin lispro analogs", LysPro insulin and humanlogs, as described in US Pat. No. 5,547,929, which is incorporated by reference herein in its entirety. (
humalog) insulin and other insulin analogues, as well as other "superinsulin analogues", in which the insulin analogues are similar to the liver-selective insulin analogues, which are more active in the liver than in adipose tissue. The ability of an object to influence serum glucose levels is substantially enhanced over conventional insulin. Preferred analogues are monomeric insulin analogues, for example insulin lispro, which are the same versatile insulin-like compounds as insulin (ie compounds used to lower blood glucose levels).

The term “amylin” refers to naturally occurring human amylin, bovine, porcine, rat, rabbit amylin, as well as, for example, pramlintide and US Pat. No. 5,686.
, 411 and US Pat. No. 5,854,215, including synthetic, anti-synthetic, or recombinant amylin or amylin agonists, such as other amylin analogs disclosed in US Pat. No. 5,854,215, both incorporated by reference in their entirety. .

The term “immunomodulatory protein” includes cytokines, chemokines, lymphokines, complement components, immune system accessories and adhesion molecules, and their receptors with human or non-human animal specificity. GM-CSF, IL-2, IL-1 are useful examples.
2, OX40, OX40L (gp34), lymphotactin, CD40, CD40L. Useful examples include interleukins such as interleukin 1-15, interferons α, β, γ, tumor necrosis factor, granulocyte macrophage colony stimulating factor (GM-CFS), macrophage colony stimulating factor (M-CSF), Granulocyte colony stimulating factor (G-CSF), eg neutrophil activating protein (NAP), macrophage chemotactic factor (MCAF), RANTES, chemokines like macrophage inflammatory peptides MIP-1a and MIP-1b, complement Components and their receptors, or for example B7.1, B7.2
, ICAM-1, 2 or 3, and accessory molecules such as cytokine receptors. OX40 and OX40-ligand (gp34) are further useful examples of immunomodulatory proteins. Immunomodulatory proteins can be used for a variety of human or non-human animal-specific purposes, and in some cases and conveniently, native proteins, variants thereof, and, for example, immunoglobulin heavy chain constant regions. Extracellular domains and other fragments that have the binding activity of fusion proteins with other polypeptide sequences such as can be representative for this purpose. Where nucleotide sequences encoding one or more immunomodulatory proteins have been inserted, those immunomodulatory proteins can include, for example, one or more cytokines, or a combination of cytokines and accessory / adhesion molecules.

As used herein, the term “interferon” or “INF” refers to a family of highly homologous species-specific proteins that inhibit viral replication and cell proliferation, as well as regulate immune responses. . Interferons are classified into three classes based on their cellular origin and antigenicity: α-interferon (white blood cells), β-interferon (fibroblasts), and γ-interferon (immunoreactive cells). Recombinant forms and analogs of each group have been developed and are commercially available. The subtypes in each group are classified based on antigenic / structural characteristics. At least 24 interferon alphas (classified into subtypes A to H) with different amino acid sequences have been identified by isolating and sequencing the DNA encoding the peptides (Viscoml, 1996 Biotherapy 10:
59-86, all or incorporated by reference). The terms "alpha-interferon,""alphainterferon,""interferonalpha,""human leukocyte interferon," and "IFN" are used interchangeably herein to describe members of that group. Both natural and recombinant alpha interferon, including consensus interferons, as described, for example, in US Pat. No. 4,897,471, which is incorporated herein by reference in its entirety, can be used in the practice of the present invention. Human leukocyte interferon prepared in such a manner comprises a mixture of human leukocyte interferons having various amino acid sequences. Purified natural human leukocyte interferons and mixtures thereof that can be used in the practice of the present invention include, but are not limited to:
Sumiferon RTM interferon α-n1; Gl available from Sumitomo (Japan)
Welfferong interferon α-n1 (Ins) available from axo-Wellcome Ltd (London, UK); and Alferon RTM interferon α-n3 available from Purdue Frederick Co., Norwalk, Conn.

The term “erythropoietin” includes synthetic, semi-synthetic, recombinant, natural, human, monkey, or other animal or microbiologically isolated mutants. Some or all of the primary structural arrangement of natural erythropoietin (ie, a contiguous sequence of amino acid residues) and one or more biological properties (eg, immunological properties and biological activity in vivo and in vitro). Applied to the polypeptide product having. The polypeptides are unique because they are the expression products of foreign DNA sequences obtained by genomic or cDNA cloning, or gene synthesis, in prokaryotic and eukaryotic hosts (eg, in cultured bacteria, yeast, and mammalian cells). Is characterized by Microbial expression products in vertebrate (eg, mammalian and avian) cells are released from human proteins or other contaminants that are associated with erythropoietin in its natural mammalian cellular environment or extracellular fluid flow, eg, plasma or urine. Is further characterized by: The products of common yeast (eg Saccharomyces cerevisiae) or prokaryotic (eg yeast) host cells are free of mammalian proteins. Depending on the host used, the polypeptides of the invention may be glycosylated with mammalian or other eukaryotic sugar chains, or may be non-glycosylated. The polypeptides of the present invention may include the first methionine amino acid residue (at amino acid position 1). The novel glycoprotein products of the present invention have one or more biological properties and, to the extent that they are of natural (eg, human) erythropoietin, are of sufficient natural (eg, human) glycan composition. (Human) Erythropoietin includes those having a primary structural arrangement in which the primary structural arrangements overlap.

The terms “heparin” and “thrombolytic agent” include, for example, heparin, low molecular weight heparin, tissue plasminogen activator (TPA), urokinase (abokinase), and other factors that control coagulation. Contains anticoagulant factors.

The terms “antiprotease” and “protease inhibitor” are used interchangeably and react with a receptor or act as an antibody, enzyme, or nucleic acid,
It applies to synthetic, semi-synthetic, recombinant, natural or non-natural, soluble or immobilized agents. They include receptors that regulate humoral immune responses, receptors that regulate cellular immune responses (eg T cell receptors), and receptors that regulate neural responses (eg glutamate receptors, glycine receptors, γ- Aminobutyric acid (GA
BA receptor). They include cytokine receptors (associated with arthritis, septic shock, transplant rejection, autoimmune diseases, and inflammatory diseases), antigens for cytotoxic T cell receptors and / or T-helper cell receptors. Includes presentation-associated major histocompatibility complex (MHC) class I and II receptors (associated with autoimmune disease), and thrombin receptors (coagulated, associated with heart disease). The list includes antibodies that recognize self-antigens, eg antibodies associated with autoimmune diseases, as well as antibodies that recognize viral (eg HIV, herpes simplex virus) and / or bacterial antigens.

The terms “hormone” and “growth factor” refer to natural, human, porcine, bovine, ovine, synthetic, semi-synthetic, or recombinant, eg, growth hormone, thyroid hormone, thyroid hormone. Releasing hormone (TRH), gonadotropin releasing hormone (GnRH), luteinizing hormone, luteinizing hormone releasing hormone (LHR)
H, eg leuprolide, deltirex, gosorelin
, Including hormones, including superagonists and antagonists such as, nafarelin, danazol, and the like. Also included are somatostatin analogs such as, for example, octrocotide (sandostatin). Other agents within its biopharmaceutical category include uterine contractions (eg oxytocin), diuresis (eg vasopressin), neutropenia (eg GCSF).
, Agents for respiratory diseases (eg, superoxide dismutase), RDS (eg, surfactant containing apoprotein as required), and the like.

The term “enzyme” includes recombinant deoxyribonucleases such as DNase (Genentech), proteases (eg serine proteases such as trypsin and thrombin), polymerases (eg RNA polymerase, DNA polymerase), reverse transcriptases and kinases. , Arthritis, osteoporosis, inflammatory disease, diabetes, allergy, organ transplant rejection, oncogene activation (eg dihydrofolate reductase)
, Enzymes involved in signal transduction, self-cycle regulation, transcription, DNA replication and repair.

The term “nucleic acid” refers to DNA or R containing natural or non-natural nucleosides.
It specifically binds to any fragment of NA or other nucleic acid or oligonucleotide through a complementary hydrogen bond, and further binds to a non-nucleic acid ligate.
Other proteinoid drugs that can bind to s). In this regard, Aptamer DN
See Bock, L., et al., Nature 355: 564-566 (1992), which reports on inhibition of thrombin-catalyzed conversion of fibrinogen to fibrin by A.

Examples of biomolecules for which lead compounds may be synthesized and selected according to the present invention include, but are not limited to, cell membrane receptor agonists and antagonists, neurotransmitters, toxins and toxins, viral epitopes, hormones, Analgesics, steroids, peptides, enzyme substrates and inhibitors, cofactors, drugs, lectins, sugars, oligonucleotides, nucleic acids, oligosaccharides, lipids, proteins, and analogs of any of these molecules.

The term “analog” has a common functional activity with the molecule of which it is considered an analog,
More commonly it refers to molecules that have common structural features.

The term “recombinant” refers to any type of cloned biopharmaceutical or genetically modified molecule expressed in a prokaryotic cell, or otherwise further processed into a second.
A combinatorial library of molecules capable of forming a combinatorial library (particularly, a molecule containing a protective group that enhances physicochemical, pharmaceutical, and clinical safety of biopharmaceuticals) is referred to.

The term “vaccine” refers to an antigen-presenting cell that is isolated or capable of activating T cells, such as activated dendritic cells, to produce a diverse cellular immune response against a given antigen. It refers to pharmaceutical compositions from which they stimulate humoral and cellular immune responses. Stimulating potent antigen presenting cells by exposing the cells to the polypeptide complex in vitro. The polypeptide complex can include a dendritic cell binding protein and a polypeptide antigen, but preferably the polypeptide antigen is either a tissue-specific tumor antigen or an oncogene product. However, other antigens such as viral antigens may be used in such combinations to effect an immune activating response. In another preferred embodiment, the dendritic cell binding protein forming part of the immunostimulatory polypeptide complex is GM-CS.
It is F. In another preferred embodiment, the polypeptide antigen is the tumor-specific antigen prostate acid phosphatase. In yet another preferred embodiment, the polypeptide antigen is any one of the oncogene product polypeptide antigens.
A polypeptide complex of a dendritic cell binding protein and a polypeptide antigen that is a linker peptide may be included. The polypeptide complex may comprise a dendritic cell binding protein covalently linked to a polypeptide antigen, such a polypeptide complex preferably being a dendritic cell binding protein, preferably GM-CSF, Formed from peptide antigens. The polypeptide antigen is preferably a tissue-specific tumor antigen, such as prostate acid phosphatase (PAP), or an oncogene product, such as Her2, p21RAS, and p53; however, such as a viral antigen. Other embodiments are also within the scope of the invention.

The term "immunoglobulin" refers to, for example, the coding or encoding of one or more gene vectors, or the conjugation of various binding moieties of nucleic acids in host defense cells, or aids in the treatment of human or animal subjects. To a host defense mechanism, such as ligating an expression vector into Drugs included in that class of polypeptides include IgG, IgE, Ig
M and IgD are mentioned, and they may be used alone or in combination with another.

For the purpose of formulations of the invention intended for inhalation into the lungs, the drug is preferably micronized and the therapeutically effective amount of the drug or a portion thereof (eg 90% or more) is particulate. Usually, the microparticles have a diameter of less than about 10 μm, preferably less than 5 μm, so that the microparticles can be inhaled into the respiratory tract and / or lungs.

The particulate drug is present in the formulation of the invention in a therapeutically effective amount, which means that the drug can be administered as a dispersion or aerosol via oral or nasal inhalation. , And preferably in a single dose or in multiple doses to produce the desired therapeutic effect. The particulate drug is usually administered as an aerosol via a conventional adapter, eg a metered dose valve, via an aerosol adapter (also known as an actuator).

The term “amount” as used herein refers to a quantity or concentration in the context. The amount of drug that makes up a therapeutically effective amount will vary based on such factors as the potency of the particular drug, the route of administration of the formulation, and the mechanical system used to administer the formulation. Naturally taking such factors into consideration, one skilled in the art can select a therapeutically effective amount for a particular drug. Generally, a therapeutically effective amount is 10 times the weight of a given fluid or high pressure gas.
0% would be about 0.001% to 5% by weight.

Suitable fluids include air, hydrocarbons such as n-butane, propane, isopentane, or propellants. Suitable propellants have sufficient vapor pressure to be useful as propellants, such as 1-6 hydrogen-containing fluorocarbons (C
HF ₂ CHF ₂ , CF ₃ CH ₂ F, CH ₂ F ₂ CH ₃ , and CF ₃ CHFCF ₃ etc.), such as 1-4 carbon perfluorocarbons (eg CF ₃ CF ₃ , CF ₃ CF ₂ CF ₃ etc.) Perfluorocarbon,
Alternatively, it is any mixture thereof. Some common suitable propellants include conventional chlorofluorocarbon (CFC) propellants, such as a mixture of propellants 11, 12, and 114. Non-CFC high pressure such as 1,1,1,2-tetrafluoroethane (high pressure gas 134a), 1,1,1,2,3,3,3-heptafluoropropane (high pressure gas 227), or mixtures thereof. Gas is preferred. The fluid or propellant is preferably present in an amount sufficient to deliver a selected quantity of drug from the aerosol enclosure to the lungs.

Choosing a suitable stabilizer. Suitable stabilizers include (1) (a) chemical formula H ₂ N
Monoamino acids -R-COOH (I), ( b) Chemical formula H ₂ N-R- (COOH) monoamino dicarboxylic acid of ₂ (II), and (c) the formula (H ₂ N) ₂ -R- COOH (III) diaminomonocarboxylic acid (where R is a linear or branched alkyl radical consisting of 1 to 22 carbon atoms: sulfide (-S-); oxide (-O-); hydroxyl (OH ); Amide (-NH); Sulfate (SO ₄ ); Chemical formula

[Chemical 1] Where X is hydrogen, halogen (F, Cl, Br, I), alkyl of 1 to 6 carbon atoms, alkoxy of 1 to 6 carbon atoms, hydroxy and nitro); and, for example, thienyl, furyl, Pyranyl, imidazolyl, pyrrolyl, tizolyl,
Heterocycles such as oxazolyl, pyridyl, and pyrimidinyl compounds may be mono- or poly-substituted with groups such as), (2) (a) hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and Acid addition salts of amino groups obtained from inorganic acids such as chloric acid and organic acids such as tartaric acid, citric acid, acetic acid, succinic acid, maleic acid, fumaric acid and oxalic acid, (b) eg glutamine etc. Carbonic acid amides; eg, oxidized and non-oxidized L-cysteinylglycine, γ-
Dipeptides such as salts and esters of L-glutamyl-L-cysteine, N-acetyl-L-cysteine-glycine; Glu-L-Glu and L-Val-Th.
Either conjugated, unconjugated, or polymeric forms of r; L-aspartyl-L-phenylalanine; muramyl dipeptide; eg L-tyrosyl-L-tyrosine, L-alanyl-L-tyrosine, L-arginyl-L-tyrosine. , L-tyrosyl-L-arginine, N-Cbz-L-Leu-L-Leu-OCH, and nutrients such as salts or esters thereof; glycyl-glycine; N-acetyl-aspartic acid-L.
Glutamic acid (NAAG); and tripeptides such as, for example, oxidized and non-oxidized γ-L-glutamyl-L-cysteinylglycine, muramyl tripeptide, (c) eg L-aspartyl-L-phenylalanine methyl ester (Aspartame®), esters of carboxylic acid groups obtained from aliphatic straight or branched chain alcohols of 1 to 6 carbon atoms, such as (3) any of the above esters, (4) any of the above. Hydrates or hemihydrates, and (5) a mixture of amino acids and amino acid derivatives.

Suitable amino acids of formula I include glycine, alanine, valine, leucine,
Isoleucine, leucylalanine, methionine, threonine, isovaline, phenylalanine, tyrosine, serine, cysteine, N-acetyl-L-cysteine, histidine, tryptophan, proline, and hydroxyproline such as trans-4-hydroxyproline. . Compounds of formula II include aspartic acid and glutamic acid, and compounds of formula III include arginine, lysine, hydroxylysine, ornithine, asparagine, and citrulline.

The fluid or aerosol formulation preferably comprises a protective colloid stabilizer in an amount effective to stabilize the formulation more than the same formulation without the stabilizer, and the agent Does not cause sedimentation, creaming and agglomeration after stirring so quickly as to prevent reproducible partitioning of the drug. A reproducible distribution can be achieved if the formulation maintains a uniform drug concentration for about 15 seconds to about 5 minutes after substantially stirring. For optimal functional and therapeutic performance of the aerosol formulation either as a dry powder or an aerosol suspension, the stabilizer is a coarse carrier (eg 20-90 μm),
Alternatively, it exists as either finely pulverized powder having a diameter of 10 μm or less.
In either case, reproducible drug administration is possible without the need to limit the patient's inspiratory maneuver. Therefore, tidal flows up to 2 liters, or 15
Excellent uniformity of the amount can be obtained at an intake air flow rate of about liter / minute to about 90 liter / minute.

The particular amount of stabilizer that makes up the effective amount depends on the particular stabilizer, the particular propellant gas, and the particular drug used in the formulation. Therefore, it is not practicable to list a particular effective amount for use with a particular formulation of the invention, but one of ordinary skill in the art can readily ascertain such an amount taking into account the factors above. However,
Generally, the stabilizer is present in an amount of about 0.0000001% to about 20%, more preferably about 0.0001% to about 1%, and most preferably about 0.001% to about 0.5% of the total formulation.
It can exist in%.

Surprisingly, it has been found that the formulations according to the invention are stable without the need to use cosolvents such as ethanol or surfactants. However, other ingredients such as conventional lubricants or surfactants, co-solvents, ethanol and the like may be present in the aerosol formulations of the present invention in suitable amounts, as will be readily identified by one of ordinary skill in the art. It doesn't matter. In this regard, reference is made to US Pat. No. 5,225,183, all of which is incorporated herein by reference.

Usually, (i) an amount of drug sufficient to provide multiple therapeutically effective doses; (ii) an amount of stabilizer sufficient to stabilize each formulation; (iii) an aerosol enclosure. A fluid or propellant gas in an amount sufficient to extrude multiple doses from; and (iv) any other optional components such as ethanol as a cosolvent: and further dispersing the components. By doing so, the preparation of the present invention can be prepared. The components can be dispersed using conventional mixers or homogenizers by stirring or by ultrasonic energy. Alternatively, the components may be dispersed using a bead mill or microfluidizer. Bulk formulations can be transferred to smaller individual aerosol bottles by using a pump-to-valve, valve-to-valve, or cold-fill method. It is not necessary that the stabilizer used in the suspension aerosol formulation be soluble in the propellant. It is possible to coat the drug particles with a suitable amount of a poorly soluble stabilizer and then add the coated particles into the formulation as described above.

To deliver the formulation of the invention, an aerosol closed container equipped with a conventional valve, preferably a metered dose valve, may be used. However, the selection of an appropriate valve device for use with an aerosol formulation will depend on the particular stabilizer and other adjuvants (if any) used, propellant gas, and the particular drug used. Neoprene and beech valve gums used in metered dose valves to deliver conventional CFS formulations have optimum delivery properties and ease of handling when used with HFC-134a or HFC-227 containing formulations. There are many cases where I cannot say that. Therefore, the diaphragm is preferably DB-218 (American
Nitrile rubber or Vist like Gasket and Rubber, Schiller Park, I11)
Certain formulations of the invention are dispensed via a valve device made of EPDM rubber such as alon ^™ (Exxon), Royalene ^™ (UniRoyal), bunaEP (Bayer). Also, FLEXOMER ^TM G
A diaphragm formed from a thermoplastic elastomeric material such as ERS 1085 NT polyolefin (Union Carbide) by extrusion, injection molding, or compression molding is suitable.

Conventional aerosol seals of coated or uncoated, anodized or non-anodized, eg aluminium, glass, stainless steel, polyethylene terephthalate, for containing the formulations according to the invention. Containers and closed containers or cans covered with epon or epoxy etc. may be used.

As with powder aerosols, conventional nebulizers can be used with the formulations of the present invention.

In order to bring about a bronchodilator effect or to treat a condition which can be treated by inhalation such as asthma, chronic obstructive pulmonary disease, the formulation of the present invention is administered to the respiratory tract and / or lung by oral inhalation. Can be transported. It is also permissible to deliver the formulations of the invention by nasal inhalation, for example to treat allergic rhinitis, rhinitis (topical), or diabetes (systemic), or to treat angina or local infections, for example. For treatment, the formulations of the invention may be delivered by topical administration (eg, buccal).

─────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁷ Identification code FI theme code (reference) A61M 13/00 A61K 37/02 (81) Designated country EP (AT, BE, CH, CY, DE, DK, ES, FI, FR, GB, GR, IE, IT, LU, MC, NL, PT, SE, TR), OA (BF, BJ, CF, CG, CI, CM, GA, GN, GW, ML, MR, NE, SN, TD, TG), AP (GH, G M, KE, LS, MW, MZ, SD, SL, SZ, TZ, UG, ZW), EA (AM, AZ, BY, KG, KZ) , MD, RU, TJ, TM), AE, AG, AL, AM, AT, AU, AZ, BA, BB, BG, BR, BY, BZ, CA, CH, CN, CR, C U, CZ, DE, DK, DM, DZ, EE, ES, FI, GB, GD, GE, GH, GM, HR, HU, ID, IL, IN, IS, JP, KE, KG, KP, KR , KZ, LC, LK, LR, LS, LT, LU, LV, MA, MD, MG, MK, MN, MW, MX, MZ, NO, NZ, PL, PT, RO, RU, SD, SE, SG, SI, SK, SL, TJ, TM, TR, TT, TZ, UA, UG, UZ, VN, YU, ZA, ZW (72) Inventor, Yapin, New Jersey, USA 08904 Highland Park South Tenth Street 284 Apartment A (72) Inventor San, John Zee, New Jersey, USA 08817 Edison Forest Haven Boulevard 1519 (72) Inventor Stefano, Simon United States of America Country New Jersey 07950 Morris Plains Meadow Bluff Road 35 F Term (reference) 4C076 AA24 BB22 BB25 DD34A DD35A DD37E DD51Q FF12 FF15 FF63 GG45 GG46 4C084 AA03 BA14 BA15 DB01 DB01 DB01 DB31 DB31 DB01 DB01 DB09 DB09 DB01 DB09 DB09 DB01 DB09 DC03 DC23 DC32 DC34 MA05 MA13 MA57 MA59 NA11

Claims (18)

[Claims] 1. A pharmaceutical formulation comprising: (a) a therapeutically effective amount of a protein or peptide drug having a molecular size of from about 1 kilodalton to about 150 kilodaltons; (b) a flow for containing said drug. A pharmaceutical formulation comprising a carrier; and (c) a stabilizer selected from amino acids, their derivatives, or mixtures thereof. 2. The drug is insulin, insulin analogue, amylin, immunomodulatory protein, interleukin, interferon, erythropoietin, heparin, thrombolytic agent, antitrypsin, antiprotease, hormone, growth factor, enzyme, nucleic acid, immunity. Globulin, antibiotics, anti-infectives, calcitonin, hematopoietic factors, vaccines, vasoactive peptides, antisense drugs, oligonucleotides, DN
A formulation according to claim 1, characterized in that it is selected from the group consisting of ase, cyclosporine, ribavirin, or any mixture thereof. 3. The drug is insulin, insulin analogue, amylin, glucagon, LH-RH, deltirex, leuprolide, gosorelin (
gosorelin), nafarelin, octreotide, somatostatin,
Calcitonin, parathyroid hormone, TRH, growth hormone releasing hormone, G-CSF, G-
SF, cytokine, rhDNase, heparin, antibiotics, albumin, ovalbumin, amiloride, DDAVP, VIP, cyclosporine, erythropoietin, interferon, IgG, IgE, IgM, IgA, IgD, interleukin, IRAP, papain, peroxidase, serachiapeptidase, The formulation according to claim 1, which is selected from the group consisting of catalase, α-1-antitrypsin, gene, vector, amiloride, rhDNase, oligonucleotide, ribavirin, or any mixture thereof. 4. The stabilizing agent is selected from the group consisting of 20 pre-existing amino acids, any mixture thereof, and derivatives thereof.
The described formulation. 5. The stabilizer is (1) a salt of oxidized and non-oxidized L-cysteinylglycine, γ-L-glutamyl-L-cysteine, N-acetyl-L-cysteine-glycine. And a dipeptide selected from the group consisting of an ester; (2) a conjugated, non-conjugated, or polymerized form of L-Gly-L-Glu and L-Val-Thr; (3) L-aspartyl-L-phenylalanine;
) Muramyl dipeptide; (5) L-tyrosyl-L-tyrosine, L-alanyl-L
-Tyrosine, L-arginyl-L-tyrosine, L-tyrosyl-L-arginine,
A nutrient selected from the group consisting of N-Cbz-L-Leu-L-Leu-OCH and salts or esters thereof; (6) glycyl-glycine; (7) N-acetyl-aspartic acid-L-glutamic acid (NAAG). ); (8) a tripeptide selected from the group consisting of oxidized and non-oxidized γ-L-glutamyl-L-cysteinylglycine or muramyl tripeptide; and (9) consisting of any mixture thereof. Formulation according to claim 1, characterized in that it is selected from the group. 6. The fluid carrier is 1,1,1,2-tetrafluoroethane, 1,1,1,2
The formulation according to claim 1, wherein the formulation is selected from the group consisting of 3,3,3,3-heptafluoropropane, or a mixture thereof. 7. The formulation according to claim 1, wherein the fluid carrier is a hydrocarbon propellant selected from the group consisting of n-butane, propane, isopentane, or a mixture thereof. 8. The formulation according to claim 1, further comprising a cosolvent. 9. The formulation of claim 8, wherein the co-solvent comprises ethanol. 10. The stabilizer is present in an amount effective to prevent settling, creaming and flocculation of the formulation for a time sufficient to result in reproducible partitioning of the drug after stirring the formulation. The formulation according to claim 1, characterized in that 11. The formulation according to claim 10, wherein the stabilizer is present in an amount ranging from about 0.0000001% to about 20% by weight, based on the total weight of the formulation. 12. A method for preparing an aerosol formulation according to claim 1, comprising: (a) (i) a sufficient amount of the drug to provide multiple therapeutically effective doses, and (ii) multiple doses. (B) the component (i) is admixed with a sufficient amount of the fluid carrier to extrude a therapeutically effective amount of , (Ii), and (iii) are dispersed: A method comprising each step. 13. The method further comprising mixing a co-solvent in the step (a) and further dispersing the components (i), (ii), (ii) together with the co-solvent in the step (b). 13. The method of claim 12 characterized. 14. A method of treating a condition treatable by oral or nasal inhalation in a human or animal, comprising the step of administering the formulation of claim 1 to said human or animal by oral or nasal inhalation. How to become. 15. The preparation according to claim 1, which is contained in an aerosol closed container with a metering valve. 16. A method of stabilizing a suspension aerosol formulation comprising a propellant gas as well as a protein or peptide drug, the method comprising: after stirring the formulation, for a time sufficient to result in a reproducible distribution of the drug. Adding to the formulation a stabilizer selected from the group consisting of suitable amino acids, their derivatives, or any mixture thereof, in an amount effective to prevent settling, creaming and aggregation of the formulation. How to become. 17. A metered dose inhaler containing a pharmaceutical aerosol formulation, comprising:
(A) a therapeutically effective amount of a protein or peptide drug; (b) propellant gas; and (c) a sufficient time after stirring the formulation to provide a reproducible distribution of the drug,
Metered dose inhaler comprising a stabilizer selected from the group consisting of suitable amino acids, their derivatives, or any mixture thereof, effective to prevent settling, creaming and aggregation of the formulation. . 18. The metered dose inhaler of claim 17, wherein the stabilizer is selected from the group consisting of 20 pre-existing amino acids, any mixture thereof, and derivatives thereof.