MXPA00006644A - Method for administering monomeric insulin analogs - Google Patents

Abstract

The claimed invention relates to a method of administering an insulin analog by inhalation, a method for treating diabetes by administering an insulin analog by inhalation, and a method for treating hyperglycemia by administering an insulin analog by inhalation.

Description

METHOD PAID TO MANAGE INSULIN MONOMERAL ANALOGUES f DESCRIPTION OF THE INVENTION This invention relates generally to methods of treating humans suffering from diabetes mellitus. More specifically, this invention relates to the pulmonary delivery of monomeric insulin analogs for systemic absorption through the lungs, in order to significantly reduce or eliminate the need for administration of monomeric insulin analogs by injection. Since the introduction of insulin in the 1920s, continuous improvements have been made in order to improve the treatment of diabetes mellitus. Major advances have been made in the purity and availability of insulin and in the various formulations with different actions with respect to time have also been developed. A non-injectable form of insulin is desirable to increase patient compliance with intense insulin therapy and a decrease in the risk of complications. Diabetes mellitus is a disease that affects approximately 6% of the world population. In addition, the population of most countries is getting older and diabetes is particularly common in elderly populations. Frequently, it is this population group that REF.120522 experience difficulty or lack of desire to administer to themselves insulin by injection. In the United States, approximately 5% of the population has diabetes and approximately one third of these diabetics self-administer one or more doses of insulin per day by subcutaneous injection. This type of intensive therapy is necessary to lower blood glucose levels. The high blood glucose concentrations, which are the result of. Low or absent concentrations of endogenous insulin, alter the normal body chemistry and can lead to failures in the microvascular system in many organs. Untreated diabetics often experience amputations and experience blindness and kidney failure. Medical treatment of the side effects of diabetes and loss of productivity due to inadequate treatment of diabetes is estimated to have an annual cost of approximately $ 40,000 million in the United States alone. The nine-year diabetes and complications management trial (DCCT), which involves 1,441 type I diabetic patients, demonstrates that maintaining blood glucose levels within tight tolerances reduces the frequency and severity of diabetes complications. Conventional insulin therapy involves only two injections per day. Intensive insulin therapy in 1 DCCT study involves three or more insulin injections each day. In this study, the incidence of side effects of diabetes is markedly reduced. For example, retinopathy is reduced by 50-76%, nephropathy by 35-56% and neuropathy by 60% in patients who use intensive therapy. Unfortunately, many diabetics do not want to undergo intensive therapy due to the discomfort associated with the many injections necessary to maintain tight control of glucose levels. This type of therapy can be psychologically and physically painful. Upon oral administration, insulin is rapidly degraded in the Gl tract and is not absorbed into the bloodstream. Therefore, many researchers have studied alternative routes to administer insulin such as the oral, rectal, transdermal and nasal. Unnow, however, these routes of administration have not resulted in effective insulin absorption. For many years it has been known that some proteins can be absorbed by the lung. In fact, the administration of insulin as an inhalation aerosol to the lung was first reported by Gaensslen in 1925. Despite the fact that many human and animal studies have shown that some insulin formulations can be absorbed through the lungs, pulmonary delivery has not received widespread acceptance as a means to effectively treat diabetes. This is partly due to the small amount of insulin which is absorbed in relation to the quantity supplied. In addition, researchers have observed a large degree of variability in the amount of insulin absorbed after pulmonary delivery of different insulin formulations at similar doses of the same formulation delivered at different times. Therefore, there is a need to provide an efficient and reliable method for delivering insulin through the lung. This need is particularly evident for patients undergoing aggressive treatment protocols using analogs of fast-acting human monomeric insulin. Efficient pulmonary delivery of fast-acting human monomeric insulin analogues can have the effect of rapidly reducing blood glucose concentrations which may be increased, such as, for example, after a meal or after a prolonged period without insulin therapy. It is evident that not all proteins can be efficiently absorbed in the lungs. There are many factors that affect whether a protein can be delivered effectively through the lungs. The absorption through the lungs depends to a large extent on the physical characteristics of the particular therapeutic protein to be delivered. Thus, although the pulmonary supply of regular human insulin has been observed, the physical differences between regular human insulin and fast-acting monomeric insulin analogs make it unclear whether these analogues can be delivered effectively through the pulmonary route. An efficient pulmonary supply of a protein depends on the ability to supply the protein in the deep pulmonary alveolar epithelium. The proteins that deposit the epithelium of the upper airways are not significantly absorbed. This is due to the overlying mucus which is about 30-40 μm thick and acts as a barrier to absorption. In addition, proteins deposited in this epithelium are cleared by mucociliary transport to the airways and then eliminated by the gastrointestinal tract. This mechanism also contributes substantially to the low absorption of some protein particles. The degree to which the proteins are not absorbed and instead are eliminated by these routes, depends on their solubility, their size as well as other characteristics less understood. It is difficult to predict whether a therapeutic protein can be transported rapidly from the lung to the blood even if the protein can be delivered successfully to the deep pulmonary alveolar epithelium. Absorption values have been calculated for some proteins delivered through the lungs, and range from 15 minutes for parathyroid hormone (fragment 1-34) to 48 hours for glycosylated l-antitrypsin. Due to the broad spectrum of peptidases which exist in the lungs, a longer absorption time increases the possibility that the protein is significantly degraded or that it is cleared by mucociliary transport before its absorption. Insulin is a peptide hormone with a molecular weight of approximately 5,800 Daltons. In the presence of zinc, human insulin self-associates in a stable, hemispheric form. It is considered that the dissociation of the stable hexamer is the limiting step in the speed with respect to the absorption of insulin from the site of subcutaneous injection into the bloodstream. However, fast-acting insulin analogs do not easily form stable hexamers. These analogs are known as monomeric insulin analogs because they are less likely to self-associate to form stable higher order complexes. This lack of self-association is due to the modifications in the amino acid sequence of human insulin that decreases the association by interrupting the formation of dimers. Unfortunately, the modifications to insulin which cause these analogs to be monomeric, also results in non-specific aggregation of monomers. This non-specific aggregation can return to insoluble and unstable analogues.

Therefore, due to the inherent instability of monomeric insulin analogues, the possibility of forming insoluble insulin analogue precipitates, the physical differences between insulin and monomeric analogs of insulins as well as the high degree of variability in the absorption of insulin. Regular human insulin delivered through the lungs, it is surprising that analogous formulations of aerosolized monomeric insulin can be delivered reproducibly and effectively through the lungs. More advantageously and unexpectedly is the discovery that, in contrast to the data obtained with regular human insulin, a change in the inhaled volume does not lead to detectable differences in either the pharmacokinetics or pharmacodynamics of the monomeric insulin analogues, particularly LysB2tfProB -human insulin Furthermore, it is surprising that LysB28ProB29-human insulin is absorbed less rapidly from the lung, after delivery subsequent to subcutaneous administration. The present invention relates to a method for administering a monomeric insulin analog comprising administering an effective amount of the monomeric insulin analogue to a patient in need thereof, by pulmonary means. The present invention also relates to a method of treating diabetes comprising administering an effective dose of a monomeric analogue of insulin to a patient in need thereof, by pulmonary means. Another aspect of the invention relates to a method for treating hyperglycemia comprising administering an effective dose of a monomeric insulin analog to a patient in need thereof, by pulmonary means. Preferably, monomeric insulin analogs are delivered by inhalation to the patient's lower airway. Monomeric insulin analogs can be supplied in. a carrier, for example a solution or suspension, or as a dry powder, using any of various devices suitable for administration by inhalation. Preferably, monomeric insulin analogs are delivered in an active particle size to reach the lower airways of the lungs. A preferred particle size for the monomeric insulin analog is less than 10 microns. An even more preferred particle size for the monomeric insulin analog is between 1 and 5 microns. Figure 1 presents a graph of the mean glucose response in hounds versus time after aerosol delivery of LysB28ProB29-human insulin. The term "insulin" as used herein, refers to mammalian insulin, such as bovine, porcine or human insulin, whose sequences and structures are known in the art. The amino acid sequence and the spatial structure of human insulin are well known. Human insulin is made up of an A chain of 21 amino acids and a B chain of 30 amino acids which are crosslinked by disulfide bonds. Appropriately cross-linked human insulin contains three disulfide bridges: one between position 7 of chain A and position 7 of chain B, the second between position 20 of chain A and position 19 of chain B and the third between positions 6 and 11 of chain A. The term "insulin analogue" means protein having an A chain and a B chain having substantially the same amino acid sequences as the , chain A and chain B of human insulin, respectively, but which differ from chain A and chain B of human insulin having one or more amino acid deletions or one or more amino acid substitutions and / or one or more additions of amino acids that do not destroy the insulin activity of the insulin analog. A type of insulin analogue the "monomeric insulin analog" is well known in the art. These are rapid-acting analogs of human insulin and include, for example, monomeric insulin analogs wherein: a) the amino acyl residue at position B28 is substituted with Asp, Lys, Leu, Val or Ala and the aminoacyl residue in position B29 is Lys or Pro; b) the aminoacyl residues at positions B28, B29 and B30 are deleted; or c) the -? or -aminoacyl residue in position B27 is deleted. A preferred monomeric insulin analog is AspB2S. A more preferred monomeric insulin analog is LysB28ProB q. Monomeric insulin analogs are described in Ch, et al., U.S. Pat. 5,514,646; Ch, et al., U.S. Patent application serial number 08 / 255,297; Brems, et al., Protein Engineering, 5: 527-533 (1992); Brange, et al. EPO publication number 214,826 (published March 18, 1987); and Brange et al., Current Opinion in Structural Biology, 1: 934-940 (1991). These descriptions are expressly incorporated herein by reference to describe monomeric insulin analogues. The insulin analogs can also have substitutions of the amino acids amidated with acid forms. For example, an Asn can be substituted with Asp or Glu. Similarly, Gln can be substituted with Asp or Glu. In particular, Asn (A18), Asn (A21) or Asp (B3) or any combination of these residues may be substituted by Asp or Glu. In addition, Gln (Al5) or GLN (B4) or both, may be substituted by either Asp or Glu. The term "conservative" refers to a compound added to a pharmaceutical formulation to act as an antimicrobial agent. A parenteral formulation must satisfy guidelines of lines for effectiveness of conservatives which are commercially available multiple use products. Among the preservatives known in the art as effective and acceptable in parenteral formulations are benzalkonium chloride, benzethonium, chlorhexidine, phenol, m-cresol, benzyl alcohol, methyl paraben, chlorobutanol, o-cresol, p-cresol, chlorocresol, phenylmercuric nitrate, thimerosal, benzoic acid and various mixtures thereof. See, for example, Wallhaser, K.-H., Develop. Biol. Standard, 24: 9-28 (Basel, S. Krager, 1974). Certain phenolic preservatives such as phenol and m-cresol are known to bind to insulin-like molecules and therefore induce conformational changes that implement physical or chemical stability, or both [Birnbaum, et al., Pharmac. Res. 14: 25-36 (1997); Rahuel-Clermont, et al., Biochemistry 36: 5837-5845 (1997)]. Preferred preservatives are m-cresol and phenol in monomeric insulin analog protein formulations used in the present invention. The term "buffer" or "pharmaceutically acceptable buffer" refers to a compound that is safe for use in insulin formulations and that has the effect of controlling the pH of the formulation at the desired pH for the formulation. Pharmaceutically acceptable buffers for controlling pH at a moderately acidic pH to a moderately basic pH include, for example, compounds such as phosphate, acetate, citrate, tris, arginine or histidine.

The term "isotonicity agent" refers to a compound that is physiologically tolerated and imparts a proper tonicity to a formulation to prevent a net flow of water through the cell membrane. Compounds such as glycerin are commonly used for such purposes at known concentrations. Other acceptable isotonicity agents include salts, for example NaCl, dextrose, mannitol and lactose. It is preferred as an isotonicity agent to glycerol at a concentration of 12 to 25 mg / ml.

Administration of Monomeric Analogs of Insulin Monomeric insulin analogs are administered by inhalation in a dose-effective manner to increase circulating insulin protein concentrations and / or to decrease circulating glucose levels. Such administration may be effective to treat disorders such as diabetes or hyperglycemia. The acquisition of effective doses of monomeric insulin analogs requires the administration of an inhaled dose of more than about 0.5 μg / kg to about 50 μg / kg of insulin analog monomeric protein, preferably from about 3 μg / kg to about 20 μg / kg, and more preferably from about 7 μg / kg to about 14 μg / kg. A therapeutically effective amount can be determined by a knowledgeable physician, who will take into consideration factors including the insulin protein concentration, blood glucose concentrations, the physical condition of the patient, the patient's lung status or the like. According to the invention, monomeric insulin analogues are supplied by inhalation to obtain a rapid absorption of these analogues. Administration by inhalation may result in a pharmacokinetics comparable to the subcutaneous administration of insulins. Inhalation of monomeric insulin analogues leads to a rapid increase in the concentration of circulating insulin after a rapid decrease in blood glucose levels. Different inhalation devices typically provide similar pharmacokinetics when comparing similar particle sizes as well as similar concentrations of lung deposition. In accordance with the invention, monomeric insulin analogs can be delivered by any of various inhalation devices known in the art for administration of a therapeutic agent by inhalation. These devices include metered dose inhalers, nebulizers, dry powder generators, sprinklers and the like. Preferably, monomeric insulin analogs are delivered by an inhaler or dry powder sprayer. There are several desirable characteristics of an inhalation device for administering monomeric insulin analogues. For example, the delivery by the inhalation device is advantageously reliable, reproducible and accurate. The inhalation device must provide small particles, for example less than about 10 μm, preferably about 1-5 μm, for good breathing capacity. Some specific examples of commercially available inhalation devices suitable for the practice of this invention are Turbohaler ™ (Astra), Ronahaler * (Glaxo), Diskus * (Glaxo), Spiros inhaler (tiffra), devices sold by Inhale Therapeutics, AERx ™ (Aradigm). ), the UltraventMB nebulizer (Mallinckrodt), the Acorn IIMR nebulizer (Marquest Medical Products), the Ventolin metered dose inhaler * (Glaxo), the Spinhaler ™ powder inhaler (Fisons) or similar. As will be recognized by those skilled in the art, the formulation of the monomeric insulin analogue protein, the amount of formulation delivered and the duration of administration of a single dose depend on the type of inhalation device used. For some aerosol delivery systems, such as nebulizers, the frequency of administration and the duration of time by which the system is activated will depend mainly on the concentration of monomeric insulin analogue in the aerosol. For example, shorter periods of administration at higher concentrations of monomeric insulin analogue protein can be used in the nebulizer solution. Devices such as metered dose inhalers can produce higher concentrations of aerosol, and can be operated for shorter periods to supply the desired amount of monomeric insulin analogue protein. Devices such as powder inhalers provide active agent until a given agent load is driven from the device. In this type of inhaler, the amount of analogous monomeric insulin protein in a given amount of powder determines the dose delivered in a single administration. The particle size of the monomeric insulin analog protein in the formulation delivered by the inhalation device is critical with respect to the ability of the protein to accumulate in the lungs, and preferably within the lower airways or alveoli. Preferably, the monomeric insulin analog is formulated so that at least about 10% of the delivered monomeric insulin analog protein is deposited in the lung, preferably about 10% to about 20% or more, the efficiency is known to be Maximum lung deposition for humans breathing through the mouth is obtained with particle sizes from approximately 2 μm to approximately 3 μm. When the particle sizes are above about 5 μm, the lung deposition decreases substantially. Particle sizes less than about 1 μm cause a decrease in lung deposition, and it becomes difficult to deliver particles with a sufficient mass to be therapeutically effective. Thus, insulin monomeric analog protein particles delivered by inhalation have a particle size preferably of less than about 10 μm, more preferably in the range of about 1 μm to about 5 μm, and much more preferably in the range from about 2 μm to about 3 μm. The monomeric insulin analog protein formulation is selected to provide the desired particle size in the effector inhalation device.

Administration of Monomeric Analogs of Insulin by a Dry Powder Inhaler Advantageously for administration as a dry powder, the analogous insulin monomeric protein is prepared in a particulate form with a particle size of less than about 10 μm, preferably from about 1 to about 5 μm, and most preferably about 2 μm to about 3 μm. The preferred particle size is effective to deliver to the alveoli of the patient's lungs. Preferably, the dry powder is constituted mainly of particles produced so that most of the particles have a size in the desired range. Advantageously, at least about 50% of the dry powder is made of particles having a diameter of less than about 10 μm. Such formulations can be obtained by spray-drying, milling or critical point condensation of a solution containing analogous monomeric insulin and other desired ingredients. Other methods also suitable for generating particles useful in the present invention are known in the art. The particles are usually separated from a dry powder formulation in a container and then transported to the patient's lung by means of a carrier air stream. Typically, in current dry powder inhalers, the force to break the solid is provided solely by the inhalation of the patient. A suitable dry powder inhaler is Turbohaler ™ manufactured by Astra (Sódertalje, Sweden). In another type of inhaler, the air flow generated by the inhalation of the patient activates a driving motor which deagglomerates the analogous monomeric particles of insulin. The inhaler Dura Spiros ™ is such a device.

Formulations of monomeric insulin analogs for administration from a dry powder inhaler typically include a finely divided dry powder containing monomeric insulin analogue protein, but the powder may also include a bulk agent, a carrier, excipient, other additive. or similar. The additives can be included in a dry powder formulation of a monomeric insulin analogue protein, for example, to dilute the powder as required for delivery from the particular powder inhaler, to facilitate the processing of the formulation, to provide advantageous properties of the powder to the formulation, to facilitate the dispersion of the powder from the inhalation device, to stabilize the formulation (for example antioxidants or buffers) to provide a flavor to the formulation, or the like. Advantageously, the additive does not adversely affect the patient's airways. The monomeric insulin analogue protein can be mixed with an additive at the molecular level or the solid formulation can include particles of the monomeric insulin analogue protein mixed with, or coated onto, particles of the additive. Typical additives include mono-, di- and polysaccharides; sugar alcohols and other polyols such as, for example, lactose, glucose, raffinose, melezitose, lactitol, maltitol, trehalose, sucrose, mannitol, starch or combinations thereof; surfactants such as sorbitols, diphosphatidylcholine or lecithin; or similar. Typically, an additive such as a bulking agent is present, in an amount effective for the purpose described above, often at about 50% to about 90% by weight of the formulation. Additional agents known in the art for formulation of a protein such as an insulin analogue protein can also be included in the formulation. The administration of a dry powder formulation of Humalog ™ which is LysB28ProB29-human insulin, by inhalation, is a preferred method of treating diabetes.

Administration of Monomeric Insulin Analogs as a Spray A spray that includes monomeric insulin analogue protein can be produced by forcing a suspension or solution of monomeric insulin analogue protein through a nozzle under pressure. The size of the nozzle and the configuration, the applied pressure and the liquid feed speed can be chosen to obtain the desired output and particle size. An electrospray can be made, for example, by an electric field in connection with a capillary or nozzle feed. Advantageously, particles of analogous insulin monomeric protein can be delivered by a spray having a particle size of less than about 10 μm, preferably in the range of about 1 μm to about 5 μm, and more preferably about 2 μm to about 3 μm. Monomeric insulin analog protein formulations suitable for use with a spray typically include monomeric insulin analog protein in an aqueous solution at a concentration of about 1 mg to about 20 mg of monomeric insulin analogue protein per ml of solution. The formulation may include agents such as an excipient, a buffer, an isotonicity agent, a preservative, a surfactant and preferably zinc. The formulation may also include an excipient or agent for stabilization of the monomeric insulin analogue protein, such as a buffer, a reducing agent, a protein in volume or a carbohydrate. Volume proteins useful for formulating monomeric insulin analog proteins include albumin, protamine or the like. Typical carbohydrates useful in formulating analogous monomeric insulin proteins include sucrose, mannitol, lactose, trehalose, glucose or the like. The monomeric insulin analogue protein formulation can also include a surfactant, which can reduce or prevent surface-induced aggregation of the monomeric insulin analogue protein caused by atomization of the solution by forming an aerosol. Various conventional surfactants can be used such as esters and polyoxyethylene fatty acid alcohols, and polyoxyethylene sorbitol fatty acid esters. The amounts generally vary between 0.001% and 4% by weight of the formulation. Especially preferred surfactants for purposes of this invention are polyoxyethylene sorbitan monooleate, polysorbate 80, polysorbate 20 or the like. Additional agents known in the art for formulation of a protein such as analogous insulin protein, can also be included in the formulation.

Administration of Monomeric Analogs of Insulin by a Nebulizer The monomeric insulin analogue protein can be administered by a nebulizer, such as a jet nebulizer or an ultrasonic nebulizer. Typically, in a jet nebulizer, a source of compressed air is used to create a high velocity jet of air through a hole. As the gas expands past the nozzle, a low pressure region is generated which extracts a monomeric insulin analogue protein solution through a capillary tube connected to a liquid reservoir. The liquid stream of the capillary tube is cut into unstable filaments and into droplets as they exit the tube, creating the aerosol. A range of configurations, flow rates and types of baffles can be used to obtain the desired performance characteristics from a given jet nebulizer. In an ultrasonic nebulizer, high frequency electrical energy is used to create vibrational mechanical energy, typically using a piezoelectric transducer. This energy is transmitted to the formulation of monomeric insulin analogue protein either directly or through a coupling fluid, creating an aerosol that includes an analogous insulin monomeric protein. Advantageously, the monomeric insulin analog protein particles supplied by a nebulizer have a particle size of less than about 10 μm, preferably in the range of about 1 μm to about 5 μm, and more preferably from about 2 μm to about 3 μm. μm. Formulations of analogous insulin monomeric protein suitable for use with a nebulizer, either jet or ultrasonic, typically include monomeric insulin analog proteins in an aqueous solution at a concentration of about 1 mg to about 20 mg of monomeric analog protein. insulin per mm of solution. The formulation may include agents such as an excipient, a buffer, an isotonicity agent, a preservative, a surfactant and, preferably, zinc. The formulation may also include an excipient or agent for stabilization of the monomeric insulin analogue protein, such as a buffer, a reducing agent, a protein in volume or a carbohydrate. Volume proteins useful for formulating monomeric insulin analog proteins include albumin, protamine or the like. Typical carbohydrates useful in the formulation of monomeric insulin analog proteins include sucrose, mannitol, lactose, trehalose, glucose or the like. The monomeric insulin analogue protein formulation may also include a surfactant, which reduces or prevents surface-induced aggregation of the monomeric insulin analogue protein caused by atomization of the solution by forming an aerosol. Various conventional surfactants can be used, such as polyoxyethylene fatty acid esters and alcohols, and polyoxyethylene sorbital fatty acid esters. The amounts generally vary between 0.001 and 4% by weight of the formulation. Especially preferred surfactants for purposes of this invention are polyoxyethylene sorbitan monooleate, polysorbate 80, polysorbate 20 or the like. Additional agents known in the art can also be included in the formulation for formulation of a protein such as an analogous insulin protein.

Administration of Monomeric Insulin Analogs by a Measured Dose Inhaler In a metered dose inhaler (MDI), a propellant, a monomeric insulin analog protein and any excipient or other additive, such as a mixture that includes a liquefied compressed gas, are contained in a container. The actuation of the metering valve releases the mixture as an aerosol which preferably contains particles in the size range of less than about 10 μm, preferably from about 1 μm to about 5 μm, and most preferably from about 2 μm to about 3 μm. The desired size of aerosol particle can be obtained by using an analogous insulin monomeric protein formulation produced by various methods known to those skilled in the art, including jet grinding, spray drying, flash point condensation or the like. Preferred metered dose inhalers include those manufactured by 3M or Glaxo and utilizing a hydrofluorocarbon propellant. Formulations of monomeric insulin analogue protein for use with a metered dose inhaler device generally include a finely divided powder containing monomeric insulin analogue protein as a suspension in a non-aqueous medium, for example, suspended in a propellant with the aid of a surfactant. The propellant may be any conventional material used for this purpose, such as a chlorofluorocarbon, a hydrochlorofluorocarbon, a hydrofluorocarbon or a hydrocarbon, which includes tri-chloro-oro-omene, dichloro-if-loromethane, diclotetrafluoroethanol and 1,1,12 -tetrafluoroethane, HFA-134a (hydrofluoroalkane-134a), HFA-227 (hydrofluoroalkane-227), or the like. Preferably, the propellant is a hydrofluorocarbon. The surfactant may be chosen to stabilize the monomeric insulin analog protein as a suspension in the propellant, to protect the active agent against chemical degradation and the like. Suitable surfactants include sorbitan triolate, soy lecithin, oleic acid or the like. In some cases aerosol solutions are preferred using solvents such as ethanol. Additional agents known in the art for formulation of a protein such as an analogous insulin protein can also be included in the formulations. A person ordinarily skilled in the art will recognize the methods of the present invention can be obtained by pulmonary administration of monomeric insulin analogs and devices not described herein.

Pharmaceutical formulations of monomeric insulin analogue protein The present invention also relates to a pharmaceutical composition or formulations that include monomeric insulin analogue and that is suitable for administration by inhalation. According to the invention, the monomeric insulin analogue protein can be used for the preparation of a formulation or medicament suitable for administration by inhalation. The invention also relates to methods for the preparation of formulations including monomeric insulin analogue protein in a form that is suitable for administration by inhalation. For example, a dry powder formulation can be made in various ways, using conventional techniques. Particles in the appropriate size range for maximum deposition in the lower respiratory tract can be made by micronization, grinding, spray drying or the like. The liquid formulation can be made by dissolving the insulin monomeric analog protein in a suitable solvent, such as water, at an appropriate pH including buffers and other excipients. A particular pharmaceutical composition for a particular monomeric insulin analogue protein to be administered via the pulmonary route is Humalog ™. Humalog ™ formulations are described by DeFelippis, U.S. Pat. No. 5,461,031; Bakaysa, et al., U.S. Patent No. 5,474,978; and Baker, et al., U.S. Pat. No. 5,504,188. These descriptions are expressly incorporated herein by reference to describe the various analogous monomeric formulations of insulin. Other formulations include solutions of sterile water alone and aqueous solutions containing low concentrations of surfactants and / or preservatives, and / or stabilizers, and / or buffers. Additional suitable formulations of monomeric insulin analogs with zinc are known to those skilled in the art. The present invention can be better understood with reference to the following examples. It is intended that these examples be representative of specific embodiments of the invention and are not intended to limit the scope of the invention.

Examples Serum pharmacokinetics of LysB2 ProB29-human insulin in hound dogs after pulmonary administration of aerosolized doses alone The aerosols of LysB28 ProB29-human insulin (LysB28 ProB29-hI), generated from solutions of LysB2f ProE29-hI in sterile water, are administered to anesthetized dogs via the pulmonary route through the endotracheal tube via an ultrasonic nebulizer. The serum concentration of immunoreactive LysB2B ProB29-hI is determined by validated radioimmunoassay methods. In this study, six hound dogs (3 males and 3 females) were used. The animals were housed in two per cage or individually in stainless steel cages with suspended mesh floors. Initially, all dogs were fed approximately 450 g of Purina 5007 certified canine diet each day. The animals were kept under fasting approximately 8 hours before dosing. After recovery from anesthesia, food and water are provided ad libitum until 48 hours after the dose. The initial daily feeding regimen was started 48 hours after the dose. At the start of the study, the animals weighed between 12.5 and 17.6 kg. Blood samples were collected at various points in time after dosing to determine the plasma concentrations of LysB28 ProB29-hI and the bioavailability of the inhaled material is determined. Dogs were chosen because they are large animals with deposition in the respiratory tract of particles similar to humans.

The pulmonary administration of LysB2? ProB29-hI results in a systemic exposure as indicated by the increased concentrations of LysB29 ProB29-hI immunoreactive in the serum of all dogs.

Table 1: serum concentrations of LysR ,, i ProR2"-hI (ng / ml) versus times after pulmonary delivery, and is shown in table 1:? S I? a Abbreviations used: h, hour; M, male; F, female; N, number of animals used in the calculations; SD, standard deviation; EEM, standard error of the mean; BLQ, below the limit of quantification (< 0.25 ng / ml). For the purpose of these calculations, BLQ is assigned with a value of zero. bN.S. = no sample. No serum sample of dog 27258 is collected before dosing (0 h). Pulmonary administration produces a rapid increase in immunoreactive insulin with peak concentrations (Tmax) occurring in most dogs approximately 5 to 20 minutes after exposure to the aerosol.

Table 2: pharmacokinetic parameters for Lys-Proht. . pulmonarily. ~ Is it added? 00 L? Abbreviations used: kg, kilogram; μg, microgram; ng, nanogram; ml, milliliter; h, hour; Craax, maximum concentration in serum; tmax, maximum serum concentration time; AUC'O, area under the curve - of the dosing time of the return to the baseline; t '"return to the baseline"; ß, terminal velocity constant; t ^, half-life; M, male, F, female; SD, standard deviation; % CV, percent coefficient of variation; N, number of animals used in the calculations.

The data indicate that pulmonary administration of aerosolized LysB26 ProB29-hI results in detectable concentrations of immunosuppressive LysB2d ProB29-hI in sera from hound dogs. LysB28 ProB29-hI is rapidly absorbed with average maximum concentrations that are obtained in less than 30 minutes. The serum concentrations of immunoreactive LysB28 ProB29-hI decrease with an average lifespan of approximately 40 minutes. No appreciable differences are observed in terms of the gender in the supply and disposal of LysB28 ProB29-hI. Blood glucose values show a decline of approximately 55% of their control values in dogs fasted after inhalation of LysB2s ProB29-hI (Figure 1). The average lung dose that is required to produce these effects is approximately 7 μg / kg measured using a gamma camera detection of Technetium99 which is used as a radiolabel in the aerosol droplets. The time required for decrease in glucose concentrations is slightly lower for inhaled LysB2S ProB29-hI than the observer after subcutaneous injections. It is noted that in relation to this date, the best method known by the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects or products to which it relates. -

Claims (50)

CLAIMS Having described the invention as above, the content of the following claims is claimed as property:

1. A method for administering a monomeric insulin analog, characterized in that it comprises administering an effective amount of a monomeric insulin analog to a patient in need thereof, by pulmonary means.

2. The method according to claim 1, characterized in that a monomeric insulin analog is delivered to a lower airway of the patient.

3. The method according to claim 2, characterized in that a monomeric insulin analog is deposited in the alveoli.

4. The method according to claim 1, characterized in that a monomeric analogue of insulin is inhaled through the mouth of the patient.

5. The method according to claim 1, characterized in that a monomeric insulin analog is administered as a pharmaceutical formulation comprising a monomeric insulin analog, in a pharmaceutically acceptable carrier.

6. The method according to claim 5, characterized in that the formulation is selected from the group consisting of a solution in an aqueous medium and a suspension in a non-aqueous medium.

7. The method according to claim 6, characterized in that the formulation is administered as an aerosol.

8. The method according to claim 5, characterized in that the formulation is in the form of a dry powder.

9. The method according to claim 5, characterized in that a monomeric insulin analog has a particle size less than about 10 microns.

10. The method according to claim 9, characterized in that a monomeric insulin analog has a particle size of about 1 to about 5 microns.

11. The method according to claim 10, characterized in that a monomeric insulin analog has a particle size of from about 2 to about 3 microns.

12. The method according to claim 1, characterized in that at least 10% of the monomeric insulin analog supplied is deposited in the lung.

13. The method according to claim 1, characterized in that a monomeric insulin analog is supplied from an inhalation device suitable for pulmonary administration and capable of depositing the insulin analogue in the lungs of the patient.

1 . The method in accordance with the claim 13, characterized in that the device is selected from the group consisting of a nebulizer, a metered dose inhaler, a dry powder inhaler and a sprinkler.

15. The method in accordance with the claim 14, characterized in that the device is a dry powder inhaler.

16. The method according to claim 14, characterized in that the drive of the device delivers about 3 μg / kg to about 20 μg / kg of a monomeric insulin analogue.

17. The method according to claim 16, characterized in that the drive of the device delivers about 7 μg / kg to about 14 μg / kg of a monomeric insulin analogue.

18. The method according to claim 1, characterized in that the monomeric insulin analog is selected from the group consisting of modified human insulins, wherein: a) the amino acyl residue at position B28 is substituted with Lys, Leu, Val or Ala and the amino acyl residue at position B29 is Lys or Pro; b) the amino acyl residues at positions B28, B29 and B30 are deleted; and c) the amino acyl residue in the position B27 is deleted.

19. The method according to claim 18, characterized in that the monomeric insulin analog is LysB2f ProB29-human insulin.

20. A method for treating diabetes, characterized in that it comprises administering an effective dose of a monomeric analogue of insulin to a patient in need thereof, by a pulmonary means.

21. The method according to claim 20, characterized in that a monomeric insulin analog is administered as a pharmaceutical formulation comprising a monomeric analogue of insulin in a pharmaceutically acceptable carrier.

22. The method according to claim 20, characterized in that the monomeric insulin analog is LysB2S ProB29-human insulin.

23. The method according to claim 20, characterized in that a monomeric insulin analog is supplied from an inhalation device suitable for pulmonary administration and capable of depositing a monomeric insulin analogue in the lungs of the patient.

24. The method according to claim 23, characterized in that the device is a sprinkler or a dry powder inhaler.

25. The method according to claim 24, characterized in that a device drive delivers about 3 μg / kg to about 20 μg / kg of a monomeric insulin analogue.

26. The method according to claim 25, characterized in that a drive of the device delivers about 7 μg / kg to about 14 μg / kg of a monomeric insulin analogue.

27. The method according to claim 20, characterized in that the monomeric insulin analog is selected from the group consisting of modified human insulins, wherein: a) the amino acyl residue at position B28 is substituted with Lys, Leu, Val or Ala and the amino acyl residue at position B29 is Lys or Pro; b) the amino acyl residues at positions B28, B29 and B30 are deleted; and c) the amino acyl residue in the position B27 is deleted.

28. A method for treating hyperglycemia, characterized in that administering an effective dose of a monomeric analogue of insulin to a patient in need of it by a pulmonary means.

29. The method according to claim 28, characterized in that a monomeric insulin analog is administered as a pharmaceutical formulation comprising the insulin analog in a pharmaceutically acceptable carrier.

30. The method according to claim 28, characterized in that the monomeric insulin analog is LysB28 ProB29 -human insulin.

31. The method according to claim 28, characterized in that a monomeric insulin analog is supplied from an inhalation device suitable for pulmonary administration and capable of depositing a monomeric analogue of insulin in the lungs of the patient.

32. The method according to claim 31, characterized in that the device is selected from the group consisting of a sprinkler and a dry powder inhaler.

33. The method according to claim 31, characterized in that a drive of the device delivers about 3 μg / kg to about 20 μg / kg of a monomeric insulin analogue.

34. The method according to claim 33, characterized in that a drive of the device delivers about 7 μg / kg to about 14 μg / kg of a monomeric insulin analogue.

35. The method according to claim 28, characterized in that the monomeric insulin analog is selected from the group consisting of modified human insulins, wherein: a) the amino acyl residue at position B28 is substituted with Lys, Leu, Val or Ala and the amino acyl residue at position B29 is Lys or Pro; b) the amino acyl residues at positions B28, B29 and B30 are deleted; and c) the amino acyl residue in the position B27 is deleted.

36. A pharmaceutical composition or formulation that includes the protein a monomeric insulin analogue and that is suitable for administration by inhalation.

37. The pharmaceutical composition or formulation according to claim 36, characterized in that the monomeric insulin analog is LysB28ProB29.

38. A pharmaceutical composition or formulation characterized in that it is adapted to carry out the claimed method according to any of claims 1 to 35.

39. The use of the protein a monomeric analog of insulin for the preparation of a suitable medication for administration by inhalation.

40. The method according to claim 39, characterized in that the monomeric insulin analog is selected from the group consisting of modified human insulins, wherein: a) the amino acyl residue at position B28 is substituted with Lys, Leu, Val or Ala and the amino acyl residue at position B29 is Lys or Pro; b) the amino acyl residues at positions B28, B29 and B30 are deleted; Y

41. The use according to claim 40, characterized in that the monomeric insulin analog is LysB28ProB29.

42. The use according to claim 39, 40 or 41, characterized in that the medicament is in the form of a solution or an aqueous medium or a suspension, or a non-aqueous medium!

43. The use according to claim 39, 40 or 41, characterized in that the medicament is in the form of an aerosol.

44. The use according to claim 39, 40 or 41, characterized in that the medicament is in the form of a dry powder.

45. The use according to claim 39, 40 or 41, characterized in that the monomeric insulin analog has a particle size less than about 10 microns.

46. The use according to claim 39, 40 or 41, characterized in that the monomeric insulin analog has a particle size of about 1 to about 5 microns.

47. The use according to claim 39, 40 or 41, characterized in that the monomeric insulin analog has a particle size of from about 2 to about 3 microns.

48. The use according to any of claims 39 to 47, characterized in that the monomeric insulin analog is administered in a dose of about 3 μg / kg and about 20 μg / kg.

49. The use according to any of claims 39 to 47, characterized in that the monomeric insulin analog is administered in a dose of approximately 7 μg / kg and approximately 14 μg / kg.

50. The use according to claim 39, characterized in that it alters a method according to any of claims 1 to 35.