AU2006200276A1 - Micellar pharmaceutical compositions for buccal and pulmonary application - Google Patents

Description

N PHARMACEUTICAL FORMULATIONS FOR BUCCAL AND PULMONARY

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o ADMINISTRATION COMPRISING AN ALKALI METAL ALKYL SULFATE AND AT LEAST THREE MICELLE-FORMING COMPOUNDS en FIELD OF THE INVENTION The present invention relates to a pharmaceutical formulation comprising a pharmaceutical agent in micellar form for oral or pulmonary application as well as to o metered dose devices containing same and methods for administering same.

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V BACKGROUND OF THE INVENTION

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0 Relatively little progress has been made over the years in reaching the target of safe and effective oral formulations for macromolecules, including peptides and proteins. Barriers to developing oral formulations for proteins and peptides include poor intrinsic permeability, lumenal and cellular enzymatic degradation, rapid clearance, and chemical instability in the gastrointestinal (GI) tract. Pharmaceutical approaches to address these barriers that have been successful with traditional small, organic drug molecules have not readily translated into effective large molecule formulations.

Various routes of administration other than injection for macromolecular pharmaceutical agents proteins and peptides) have been explored with little or no success. Oral and nasal cavities have been of particular interest. The ability of molecules to permeate the oral mucosae appears to be related to molecular size, lipid solubility and peptide protein ionization. Molecules less than 1000 daltons appear to cross oral mucosae rapidly. As molecular size increases, the permeability of the molecule decreases rapidly. Lipid soluble compounds are more permeable than non-lipid soluble molecules. Maximum absorption occurs when molecules are un-ionized or neutral in electrical charges.

Charged molecules, therefore, present the biggest challenges to absorption through the oral mucosae.

Most proteinic drug molecules are extremely large molecules with molecular weights exceeding 6000 daltons. In addition to being large, these molecules typically have very poor lipid solubility, and are often practically impermeable. Substances that facilitate the absorption or transport of large molecules >1000 daltons) across biological Smembranes are referred to in the art as "enhancers" or "absorption aids." These

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O compounds generally include chelators, bile salts, fatty acids, synthetic hydrophilic and hydrophobic compounds, and biodegradable polymeric compounds. Many enhancers lack a satisfactory safety profile respecting irritation, lowering of the barrier function, and Cf impairment of the mucocilliary clearance protective mechanism.

ISome enhancers, especially those related to bile salts, and some protein solubilizing agents give an extremely bitter and unpleasant taste. This makes their use almost Oimpossible for human consumption on a daily basis. Several approaches attempting to Iaddress the taste problem relating to the bile salt-based delivery systems include patches o for buccal mucosa, bilayer tablets, controlled release tablets, use of protease inhibitors, and various polymer matrices. These technologies fail to deliver macromolecular pharmaceutical agents in the required therapeutic concentrations, however. Further, the film patch devices result in severe tissue damage in the mouth. Other attempts to deliver large molecules via the oral, nasal, rectal, and vaginal routes using single bile acids or enhancing agents in combination with protease inhibitors and biodegradable polymeric materials similarly failed to achieve therapeutic levels of macromolecular pharmaceutical agents in the patient. Single enhancing agents fail to loosen tight cellular junctions in the oral, nasal, rectal and vaginal cavities for the time needed to permit passage of large molecules through the mucosal membranes without further degradation. These problems make it impractical to use many systems. Accordingly, there remains a need for improved therapeutic formulations, particularly those comprising macromolecules and particularly those useful for buccal and pulmonary application. Methods for manufacture and use of such formulations are also needed.

SUMMARY OF THE INVENTION The present invention addresses the above need by providing a pharmaceutical formulation comprising a macromolecular pharmaceutical agent, an alkali metal alkyl sulfate, and at least three additional micelle-forming compounds, in a suitable solvent.

The agent can be one or more proteins, peptides, hormones, vaccines or drugs. The molecular weight of the macromolecular pharmaceutical agent preferably ranges between about 1,000 and 2,000,000 daltons. The agent is presented in mixed micellar form, with a I micelle size of greater than 6 pm, but may be as small as approximately one to 0 o nanometers (nm).

e As used herein the term "mixed micelles" refers to at least two different types of micelles en each of which has been formed using different micelle forming compounds. For example, the present formulations comprise a mix of at least four different types of Imicelles--micelles formed between the pharmaceutical agent and alkali metal alkyl sulfate, and micelles formed between the pharmaceutical agent and at least three different 0 oadditional micelle forming compounds as disclosed herein. It will be understood that Ieach individual micelle can be formed from more than one micelle-forming compound as o well. The mixed micelles of the present invention tend to be smaller than the pores of the membranes in the oral cavity. It is therefore believed that the extremely small size of the present mixed micelles helps the encapsulated macromolecules penetrate efficiently through the oral mucosae. Thus, the present formulations offer increased bioavailability of active drug, particularly across oral mucosae, when compared with pharmaceutical preparations known in the art.

Methods for making and using the present pharmaceutical formulations are also within the scope of the present invention. For example, the present invention may be directed to a method for enhancing the rate of absorption of a macromolecular pharmaceutical agent comprising administering a formulation comprising the agent in combination with an alkali metal alkyl sulfate and at least three micelle-forming compounds. Such a method is particularly effective when the formulation is administered to the buccal region.

Preferably they are delivered through metered dose spray devices such as metered dose dispensers (aerosol and non-aerosol). Metered dose aerosol dispensers employ propellants which are believed to provide improvements in penetration and absorption of mixed micellar formulations. Metered dose inhalers are known and are a popular pulmonary drug delivery form for some drugs.

One of the other benefits of using a metered dose dispenser, including an atomizer or inhaler, is that the potential for contamination is minimized because the devices are selfcontained.

N0 These and other aspects of the invention will be apparent from the following disclosure

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O and appended claims.

SDETAILED DESCRIPTION OF THE INVENTION Cl The present invention is directed to a pharmaceutical formulation comprising: an effective amount of a macromolecular pharmaceutical agent; an alkali metal alkyl sulfate; at least three micelle-forming compounds selected from the group comprising lecithin, Cl| 0 hyaluronic acid, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic 0 Cl acid, linoleic acid, linolenic acid, monoolein, monooleates, monolaurates, borage oil, 0 evening primrose oil, menthol, trihydroxy oxo cholanyl glycine, glycerin, polyglycerin, Cl lysine, polylysine, triolein, polyoxyethylene ethers, polidocanol alkyl ethers, chenodeoxycholate, deoxycholate, pharmaceutically acceptable salts thereof, analogues thereof and mixtures or combinations thereof; and a suitable solvent.

The alkali metal alkyl sulfate concentration is between about 0.1 and 20 wt./wt. or about 1 and 20 wt./wt. of the total formulation, each micelle-forming compound concentration is between about 0.1 and 20 wt./wt. or about 1 and 20 wt./wt. of the total formulation, and the total concentration of the alkali metal alkyl sulfate and the micelle-forming compounds together is less than 50 wt./wt. of the formulation.

As used herein, the term "macromolecular" refers to pharmaceutical agents having a molecular weight greater than about 1000 or 2000 daltons. Preferably the macromolecular pharmaceutical agents of the present invention have a molecular weight less than about 2,000,000 daltons, though larger molecules are also contemplated.

The term "pharmaceutical agent" as used herein covers a wide spectrum of agents, and can include agents used for both human and veterinary applications including but not limited to treatment and study. The term broadly includes proteins, peptides, hormones, vaccines and drugs.

Preferred pharmaceutical agents include insulin, heparin, low molecular weight heparin (molecular weight less than about 5000 daltons), hirulog, hirugen, huridin, interferons, cytokines, mono and polyclonal antibodies, immunoglobins, chemotherapeutic agents, vaccines, glycoproteins, bacterial toxoids, hormones, calcitonins, glucagon like peptides N0 (GLP-1), large molecule antibiotics greater than about 1000 daltons), protein based

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0 thrombolytic compounds, platelet inhibitors, DNA, RNA, gene therapeutics, antisense Soligonucleotides, opioids, narcotics, hypnotics, steroids and pain killers.

Ce Hormones which may be included in the present formulations include but are not limited to thyroids, androgens, estrogens, prostaglandins, somatotropins, gonadotropins, 0 erythropoetin, interferons, steroids and cytokines. Cytokines are small proteins with the Sproperties of locally acting hormones and include, but are not limited to, various forms of Sinterleukin (IL) and growth factors including various forms of transforming growth factor O (TOP), fibroblast growth factor (FGF) and insulin-like growth factor (IGF). Vaccines 0 which may be used in the formulations according to the present invention include bacterial and viral vaccines such as vaccines for hepatitis, influenza, tuberculosis, canary pox, chicken pox, measles, mumps, rubella, pneumonia, BCG, HIV and AIDS; bacterial toxoids include but are not limited to diphtheria, tetanus, Pseudomonas sp. and Mycobacterium tuberculosis. Examples of drugs, more specifically cardiovascular or thrombolytic agents, include heparin, hirugen, hirulos and hirudin. Macromolecular pharmaceutical agents included in the present invention further include monoclonal antibodies, polyclonal antibodies and immunoglobins. This list is not intended to be exhaustive.

A preferred macromolecular pharmaceutical agent according to the present invention is insulin. "Insulin" as used herein encompasses naturally extracted human insulin, insulin extracted from bovine, porcine or other mammalian sources, recombinantly produced human, bovine, porcine or other mammalian insulin, insulin analogues, insulin derivatives, and mixtures of any of these insulin products. The term further encompasses the insulin polypeptide in either its substantially purified form, or in its commercially available form in which additional excipients are added. Various forms of insulin are widely commercially available. An "insulin analogue" encompasses any of the insulins defined above wherein one or more of the amino acids within the polypeptide chain has been replaced with an alternative amino acid, wherein one or more of the amino acids has been deleted, or wherein one or more amino acids is added. "Derivatives" of insulin refers to insulin or analogues thereof wherein at least one organic substituent is bound to one or more of the amino acids in the insulin chain.

I The macromolecular pharmaceutical agent exists in micellar form in the present 0 O pharmaceutical formulations. As will be appreciated by those skilled in the art, a micelle is a colloidal aggregate of amphipathic molecules in which the polar hydrophilic portions of the molecule extend outwardly while the non-polar hydrophobic portions extend cinwardly. As discussed below, various combinations of micelle-forming compounds are utilized in order to achieve the present formulation. It is believed that the presence of the Imicelles significantly aids in the absorption of the macromolecular pharmaceutical agent oboth because of their enhanced absorption ability, and also because of their size. In 0addition, encapsulating pharmaceutical agents in micelles protects the agents from rapid o degradation in the GI environment.

0 It is believed that the mixed micelles of the present invention encapsulate molecules with a high degree of efficiency encapsulation). In general the size of the micelle particles in the mixed micellar formulation is about I to 10 nm or less, and preferably from 1 to 5 nm. Such mixed micelles tend to be smaller than the pores of the membranes in the oral cavity or the GI tract. It is therefore believed that the extremely small size of mixed micelles helps the encapsulated molecules penetrate efficiently through the mucosal membranes of the oral cavity.

The shape of the micelle can vary and can be, for example, prolate, oblate or spherical; spherical micelles are most typical.

An effective amount of the macromolecular pharmaceutical agent should be included in the present formulation. As used herein, the term "effective amount" refers to that amount of the pharmaceutical agent needed to bring about the desired result, such as obtaining the intended treatment or prevention of a disorder in a patient, or regulating a physiological condition in a patient. Such an amount will therefore be understood as having a therapeutic and/or prophylactic effect in a patient. As used herein, the term "patient" refers to members of the animal kingdom, including but not limited to humans. It will be appreciated that the effective amount will vary depending on the particular agent used, the parameters determined for the agent, the nature and severity of the disorder being treated, the patient being treated, and the route of administration. The determination of what constitutes an effective amount is well within the skill of one practicing in the art.

Typically, the present formulations will contain pharmaceutical agents in a concentration IN between about 0.1 and 20 wt./wt. or about 1 and 20 wt./wt. or about 1 and 0 Swt./wt. of the total formulation.

Cl SThe specific concentrations of the essential ingredients can be determined by relatively c€n straightforward experimentation. It will be understood that the amounts of certain ingredients may need to be limited in order to avoid formulations which produce foam when sprayed rather than forming a fine spray.

SThe amount of pharmaceutical agent in the formulations of this invention is typically a C-I quantity that provides an effective amount of the pharmaceutical agent to produce the 0physiological activity (therapeutic plasma level) for which the pharmaceutical agent is Cl being administered. In consideration of the fact that the bioavailability of any active substance can never be 100%, that is to say the administered dose of the active drug is not completely absorbed, it is preferable to incorporate a slightly larger amount than the desired dosage. For absorption through the oral cavities, it is often desirable to increase, e.g. double or triple, the dosage which is normally required through injection or administration through the gastrointestinal tract.

Any alkali metal alkyl sulfate can be used in the present formulations, provided compatibility problems do not arise. Preferably, the alkyl is a C8 to C22 alkyl, more preferably lauryl (C12). Any alkali metal can be utilized, with sodium being preferred.

The alkali metal alkyl sulfate is generally present in a concentration of between about 0.1 and 20 wt./wt. or about 1 and 20 wt./wt. or less than about 5 wt./wt. or from about 2 to 5 wt./wt. of the total formulation.

The formulations of the present invention further comprise at least three micelle-forming compounds selected from the group comprising lecithin, hyaluronic acid, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic acid, linoleic acid, linolenic acid, monoolein, monooleates, monolaurates, borage oil, evening primrose oil, menthol, trihydroxy oxocholanyl glycine, glycerin, polyglycerin, lysine, polylysine, triolein, polyoxyethylene ethers, polidocanol alkyl ethers, chenodeoxycholate and deoxycholate.

Pharmaceutically acceptable salts and analogues of any of these compounds are also within the present scope as are mixtures or combinations of any of these compounds.

Each of the three or more micelle-forming compounds listed above is present in the I0 formulations in a concentration of between about 0.1 and 20 wt./wt. or about 1 and

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0 wt./wt. or less than about 5 wt./wt. of the total formulation.

For delivery of the present macromolecular pharmaceutical agents, particularly insulin, Ce use of three or more micelle-forming compounds achieves a cumulative effect in which the amount of pharmaceutical agent that can be delivered is greatly increased as compared to when only one or two micelle-forming compounds are used. Use of three or C more micelle-forming compounds also enhances the stability of the pharmaceutical agent Sformulations.

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O The alkali metal alkyl sulfate functions as a micelle forming agent, and is added to the 0 Cl formulation in addition to the three or more other micelle-forming compounds listed herein. The total concentration of alkali metal alkyl sulfate and the three or more additional micelle-forming compounds together is less than 50 wt./wt. of the formulation.

It will be appreciated that several of the micelle-forming compounds are generally described as fatty acids, bile acids, or salts thereof. The best micelle-forming compounds to use may vary depending on the pharmaceutical agent used and can be readily determined by one skilled in the art. In general, bile salts are especially suitable for use with hydrophilic drugs and fatty acid salts are especially suitable for use with lipophilic drugs. Because the present invention uses relatively low concentrations of bile salts, problems of toxicity associated with the use of these salts is minimized, if not avoided.

The lecithin can be saturated or unsaturated, and is preferably selected from the group comprising phosphatidylcholine, phosphatidylserine, sphingomyelin, phosphatidylethanolamine, cephalin, and lysolecithin.

In one embodiment, at least one of the micelle forming compounds is selected from the group consisting of hyaluronic acid, pharmaceutically acceptable salts of hyaluronic acid, polidocanol alkyl ethers, trihydroxy oxo cholanyl glycine, polyoxyethylene ethers, triolein and mixtures thereof.

Preferred salts of hyaluronic acid are alkali metal hyaluronates, especially sodium hyaluronate, alkaline earth hyaluronates, and aluminum hyaluronate. When using I0 hyaluronic acid or pharmaceutically acceptable salts thereof in the present formulations, 0 the concentration of such ingredient may be between about 0.1 and 5 wt./wt. or about S1 and 5 wt./wt. or less than about 3.5 wt./wt. or between about 1.5 and 3.5 wt./wt.

i of the total formulation.

The above-described components of the present formulation are contained in a suitable \D solvent. The term "suitable solvent" is used herein to refer to any solvent in which the CI components of the present invention can be solubilized, in which compatibility problems o do not arise, and which can be administered to a patient. Any suitable aqueous or I\ nonaqueous solvent can be used. A particular preferred solvent is water. Other suitable 0 solvents include alcohol solutions, especially ethanol. Alcohol should be used at concentrations that will avoid precipitation of the components of the present formulations.

Enough of the solvent should be added so that the total of all of the components in the formulation is 100 wt./wt. solvent to q.s. Typically, some portion of the solvent will be used initially to solubilize the pharmaceutical agent prior to the addition of the micelle-forming compounds.

The present formulations optionally contain a stabilizer and/or a preservative. Phenolic compounds are particularly suited for this purpose as they not only stabilize the formulation, but they also protect against bacterial growth and help absorption of the formulation. A phenolic compound will be understood as referring to a compound having one or more hydroxy groups attached directly to a benzene ring. Preferred phenolic compounds according to the present invention include phenol and methyl phenol (also known as m-cresol), and mixtures thereof. The phenolic compound may be present in a concentration of from 1 to 10 wt./wt. of the total formulation.

Particularly suitable micelle-forming compound combinations include: sodium hyaluronate, monoolein and saturated phospholipid; saturated phospholipid, monoolein and glycolic acid; sodium hyaluronate, polyoxyethylene ether and lecithin; polyoxyethylene ether, trihydroxy oxocholanyl glycine and lecithin; polidocanol 9 lauryl ether, polylysine and triolein; saturated phospholipid, polyoxyethylene ether and glycolic acid; trihydroxy oxocholanyl glycine, lecithin and chenodeoxycholate; S0 trihydroxy oxocholanyl glycine, deoxycholate and glycerin; S(i) polidocanol 10 lauryl ether, sodium oxocholanyl glycine and lecithin; S(j) polidocanol 10 lauryl ether, phosphatidyl choline and oleic acid; polidocanol 10 lauryl ether, sodium hyaluronate and lecithin; and Cf, polidocanol 20 lauryl ether, evening primrose oil and lecithin.

\D The formulations of the present invention can further comprise one or more of the following: inorganic salts; antioxidants; protease inhibitors; and isotonic agents. The Samount of any of these optional ingredients to use in the present formulations can be \D determined by one skilled in the art.

0 Cl Useful inorganic salts include salts which open channels in the GI tract and which may provide additional stimulation to release insulin. Non-limiting examples of useful inorganic salts are sodium, potassium, calcium and zinc salts, especially sodium chloride, potassium chloride, calcium chloride, zinc chloride and sodium bicarbonate.

It will be recognized by those skilled in the art that for many pharmaceutical formulations it is usual to add at least one antioxidant to prevent degradation and oxidation of the pharmaceutically active ingredients. The antioxidant can be selected from the group comprising tocopherol, deteroxime mesylate, methyl paraben, ethyl paraben, ascorbic acid and mixtures thereof, as well as other antioxidants known in the pharmaceutical arts.

A preferred antioxidant is tocopherol. The parabens will also provide preservation to the formulation.

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Protease inhibitors serve to inhibit degradation of the pharmaceutical agent by the action of proteolytic enzymes. When used, protease inhibitors are preferably in a concentration of between about 0.1 and 3 wt./wt. or about 1 and 3 wt./wt. of the formulation. Any material that can inhibit proteolytic activity can be used, absent compatibility problems.

Examples include but are not limited to bacitracin and bacitracin derivatives such as bacitracin methylene disalicylates, soybean trypsin, and aprotinin. Bacitracin and its derivatives are preferably used in a concentration of between 1.5 and 2 wt./wt. of the total formulation, while soyabean trypsin and aprotinin are preferably used in a concentration of between about 1 and 2 wt./wt. of the total formulation.

IAn isotonic agent such as glycerin or dibasic sodium phosphate may also be added after

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Oformation of the mixed micellar formulation. The isotonic agent serves to keep the micelles in solution. When glycerin is used as one of the micelle-forming compounds it will also function as an isotonic agent. When dibasic sodium phosphate is used it will also cserve to inhibit bacterial growth.

IIt will also be understood by those skilled in the art that colorants, flavouring agents and non-therapeutic amounts of other compounds may be included in the formulation. Typical O flavouring agents are menthol, sorbitol and fruit flavours. When menthol is used as one I of the micelle-forming compounds, therefore, it will also impart flavor to the formulation.

N The pH of the present pharmaceutical formulation should typically be in the range of 5 to 8, more preferably 6 to 7. Hydrochloric acid or sodium hydroxide can be utilized to adjust the pH of the formulation as needed.

The formulations of the present invention may be stored at room temperature or at cold temperature. Storage of proteinic drugs is preferable at a cold temperature to prevent degradation of the drugs and to extend their shelf life.

The present invention, therefore, provides a pharmaceutical formulation in which a macromolecular pharmaceutical agent is encapsulated in mixed micelles formed by a combination of micelle-forming agents. The formulation can be delivered through buccal or pulmonary means, with buccal being preferred. Both the oral and nasal membranes offer delivery advantages, in that drugs administered through these membranes have a rapid drug absorption and a rapid onset of action, provide therapeutic plasma levels, avoid the first pass effect of hepatic metabolism, and avoid exposure of the drug to the hostile GI environment. An additional advantage is the easy access to membrane sites, so that the drug can be applied, localized and removed easily.

Oral routes of administration may be particularly advantageous. The sublingual mucosa includes the membrane of the ventral surface of the tongue and the floor of the mouth, and the buccal mucosa is the lining of the cheeks. The sublingual and buccal mucosae are relatively permeable, allowing for the rapid absorption and acceptable bioavailability of many drugs. Further, the buccal and sublingual mucosae are convenient, non-evasive and easily accessible. In comparison to the GI tract and other organs, the buccal environment

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N0 has lower enzymatic activity and a neutral pH that allows for a longer effective life of the

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O drug in vivo. The sublingual mucosa and buccal mucosa are collectively referred to herein Sas the "oral mucosae".

Ce Preferably, the present formulations are delivered through metered dose dispensers or spray devices. Metered dose dispensers are known and are a popular pulmonary drug delivery form for some drugs. One benefit of using metered dose dispensers is the ability C to deliver a precise amount of medication with each application, and another is that the 0 o potential for contamination is minimized because the devices are self-contained. The I\ metered dose dispensers may be metered dose aerosol dispensers containing a propellant.

0 Ci It is believed that improvements in penetration and absorption of the present mixed micellar formulations can be achieved by administering the present formulations with a propellant selected from the group comprising C1-C2 dialkyl ether, butanes, fluorocarbon propellant, hydrogen-containing fluorocarbon propellant, chlorofluorocarbon propellant, hydrogen-containing chlorofluorocarbon propellant, and mixtures thereof. The propellants may be tetrafluoroethane, heptafluoroethane, heptafluoropropane, dimethylfluoropropane, tetrafluoropropane, butane, n-butane, isobutane, dimethyl ether and other non-CFC and CFC propellants. The preferred propellants are hydrogencontaining chlorofluorocarbons, hydrogen-containing fluorocarbons, dimethyl ether and diethyl ether. Even more preferred is HFA-134 a (1,1,1,2-tetrafluoroethane).

Preferably, the ratio of pharmaceutical agent to propellant is from 5:95 to 25:75.

The present invention also provides a process for making the pharmaceutical formulation of the present invention. The present formulations may be prepared by mixing a solution of the macromolecular pharmaceutical agent, the alkali metal alkyl sulfate, at least three micelle-forming compounds, and optionally the stabilizer and other additives. The pharmaceutical agent should be added in an amount effective for the desired purpose. The micelle-forming compounds may be added concurrently or sequentially. Mixed micelles will form with substantially any kind of mixing of the ingredients but vigorous mixing is preferred in order to provide micelles of about 10 nanometers or less in size. The pharmaceutical agents, solvents, alkali metal alkyl sulfates, micelle-forming compounds and optional additives as described above for the present formulations are all suitable for use in the present process.

IND In one method a first micellar formulation is prepared by mixing a solution comprising

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O the pharmaceutically active agent with at least the alkali metal alkyl sulfate to form the first micellar formulation. The first micellar formulation is then mixed with at least three additional micelle-forming compounds to form a mixed micellar formulation. In another method, a first micellar formulation is prepared by mixing a solution containing the pharmaceutically active agent, the alkali metal alkyl sulfate and at least one additional ID micelle-forming compound; to the formulation is then added the remaining micelleforming compounds, with vigorous mixing. The alkali metal alkyl sulfate and three or more micelle-forming compounds should not be added to the pharmaceutical agent o solution all at once.

The stabilizer, preferably phenol and/or m-cresol, may be added to the mixed micellar formulation to stabilize the formulation and protect against bacterial growth.

Alternatively, the stabilizer may be added at the same time as any of the micelle-forming ingredients. An isotonic agent may also be added after formation of the mixed micellar formulation. Similarly, any of the other optional additives as described above can be added at this time. The formulation can then be put into an aerosol dispenser and the dispenser charged with propellant, if administration by this route is desired. The propellant, which is under pressure, is in liquid form in the dispenser. When the formulation of the present invention is in a dispenser, the aqueous phase may be separated from the propellant phase. Preferably, however, the ratios of the ingredients are adjusted by simple experimentation so that the aqueous and propellant phases become one, i.e., there is one phase. If there are two phases, it may be necessary to shake the dispenser prior to dispensing a portion of the contents, such as through a metered valve. The dispensed dose of pharmaceutical agent is propelled from the metered valve in a fine spray.

One specific embodiment of the present processes provides for making the present pharmaceutical formulations by: a) mixing a macromolecular pharmaceutical agent in a suitable solvent with an alkali metal alkyl sulfate, and adding to the mixture at least three micelle-forming compounds selected from the group comprising lecithin, hyaluronic acid, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic acid, linoleic acid, linolenic acid, monoolein, I0 monooleates, monolaurates, borage oil, evening primrose oil, menthol, trihydroxy oxo

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0 cholanyl glycine, glycerin, polyglycerin, lysine, polylysine, triolein, polyoxyethylene F1 ethers, polidocanol alkyl ethers, chenodeoxycholate, deoxycholate, pharmaceutically acceptable salts thereof, analogues thereof, and mixtures or combinations thereof, to form a mixed micellar macromolecular pharmaceutical agent formulation.

Each of the micelle-forming compounds, including the alkali metal alkyl sulfate, is in a Ci concentration of from 0.1 to 20 wt./wt. or from about 1 to 20 wt./wt. of the total Sformulation, with the total being less than 50 wt./wt. of the total formulation.

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O The method can further comprise the step of adding a stabilizer such as a phenolic 0 C] compound selected from the group comprising phenol, m-cresol and mixtures thereof; the addition of the stabilizer can be either before, during, or after the addition of the alkali metal alkyl sulfate, or before, during or after the addition of the micelle-forming compounds.

The method can further comprise the step of placing the formulation into an aerosol dispenser and charging the dispenser with a propellant.

In another specific embodiment, the process comprises: a) mixing a macromolecular pharmaceutical agent in a suitable solvent with an alkali metal alkyl sulfate, and at least one micelle-forming compound selected from the group comprising lecithin, hyaluronic acid, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic acid, linoleic acid, linolenic acid, monoolein, monooleates, monolaurates, borage oil, evening primrose oil, menthol, trihydroxy oxo cholanyl glycine, glycerin, polyglycerin, lysine, polylysine, triolein, polydocano alkyl ethers, polidocanol alkyl ethers, chenodeoxycholate, deoxycholate, pharmaceutically acceptable salts thereof, analogues thereof, and mixtures or combinations thereof, to form a first mixed micellar macromolecular pharmaceutical agent formulation; and b) adding at least two micelle-forming compounds to the first formulation that are different from that added in step a) but selected from the same group.

Again, during or after step a stabilizer as described above can be added to the formulation. Mixing can be vigorous or not. Vigorous mixing may be accomplished by I using high-speed stirrers, such as magnetic stirrers, propeller stirrers, or sonicators, and is

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o preferred.

The present invention also provides a metered dose aerosol dispenser with the n formulation of the present invention and a propellant contained therein, in which a solution containing the macromolecular pharmaceutical agent and the propellant are in a Isingle phase or in a separate phase. When in a separate phase, the aerosol dispenser must I be shaken in order to intermix the aqueous and propellant phases prior to use.

The present invention also provides a method for administering the pharmaceutical o formulations of the present invention, by spraying the intermixed formulation into the mouth with a metered dose spray device. Application can be to the buccal cavity by spraying into the cavity, without inhalation. It may be necessary or desirable to shake the dispenser prior to spraying the present pharmaceutical formulation and propellant into the buccal cavity. The plasma levels and blood glucose levels when orally administering the present insulin-containing formulations are comparable to those achieved when insulin is injected; the present methods offer significant improvements in the quality of life over injection including pain-free and needle-free therapy and improved convenience.

In the case of insulin, which is intended for administration through the mouth cavity, a first micellar solution may be made by adding water or other solvent, and then hydrochloric acid (typically 5M) to powdered insulin, and stirring until the powder is dissolved and a clear solution is obtained. The solution can then neutralized with sodium hydroxide. A sodium alkyl sulfate may be added to the neutralized solution with low speed stirring, either alone or with at least one micelle forming compound. A typical concentration of sodium lauryl sulfate, as the sodium alkyl sulfate, in the aqueous solution is less than about 20 wt./wt. or less than about 4 or 5 wt./wt. or from about 2 or 3 wt./wt. of the solution. Typically, insulin is present in the micellar solution in an amount which will give a concentration of about 0.1 to 20 wt./wt. or about I to wt./wt. of the final formulation.

The solution so formed may then be mixed vigorously, such as by sonication or high speed stirring, to form a micelle solution. Other micelle forming compounds, as described above, may then be added. The mixing may be done with a high-speed mixer or sonicator to ensure uniform micelle particle size distribution within the formulation.

I In a preferred embodiment, after forming the present micellar pharmaceutical 0 0 formulations, the phenol and/or m-cresol is added. As indicated above, other ingredients, such as isotonic agents, flavoring agents, anti-oxidants, salts, protease inhibitors or other pharmaceutically acceptable compounds may also be added to an aerosol dispenser. The formulation can be placed in an aerosol dispenser, and the dispenser charged with propellant in a known manner.

cThe specific concentrations of the above ingredients can be determined by one skilled in 0 o the art based upon the general guidelines provided herein. It will be understood that the I amounts of certain ingredients may need to be limited in order to avoid formulations O which produce foam when sprayed rather than forming a fine spray. For absorption through the oral cavities, it is often desirable to increase, such as by doubling or tripling, the dosage of pharmaceutical agent which is normally required through injection or administration through the gastrointestinal tract.

The desired size of aerosol droplets which are sprayed from the aerosol dispenser will depend, in part, on where the pharmaceutical is to be deposited. For example, for deposition in the lungs, particle sizes of less than about 5 pm are preferred whereas for absorption in the buccal cavity of the mouth, particle sizes of about 5 or 6-10 pm are preferred.

The present invention is also directed to a method for enhancing the rate of absorption of a macromolecular pharmaceutical agent comprising administering a formulation comprising said agent in conjunction with an alkali metal alkyl sulfate and at least three of the micelle-forming compounds described above. Preferably, this method is carried out by administering directly to the buccal region of the patient.

Administration of the formulation into the buccal cavity, according to any of the present methods, is by spraying the formulation into the mouth, without inhalation, so that the droplets stay in the mouth rather than being drawn into the lungs.

EXAMPLES

The following examples are intended to illustrate the invention, and should not be considered as limiting the invention in any way.

IN Example 1 c About 1000 mg of powdered insulin were placed in a glass beaker equipped with a stirrer.

c Ten ml of distilled water were added and the solution was stirred at low speed. To this eCn solution was added 5M HC1 (pH 2) solution dropwise until the insulin was solubilized completely. This solution was then neutralized, while stirring slowly, with 5M NaOH \O solution dropwise until the pH was between about 7 and 8. To this solution was added C- mg sodium lauryl sulfate, 36 mg deoxycholate, 50 mg trihydroxy oxocholanyl glycine 0 S(sodium glycocholate) and 20 mg dibasic Na phosphate; the compounds were dissolved I\ completely. 250 mg glycerin were then added while stirring at high speed, i.e. 2000 rpm.

0 o The solution was stirred for 30 minutes and then stored at 10 0 C. To this mixture 40 mg m-cresol and 40 mg phenol were added. Chenodeoxycholate or polyoxyethylene ethers can be used in place of the deoxycholate.

The solution was pipetted (1 ml/vial) into 10 ml capacity glass vials. The vials were charged with HFA-134a propellant and stored at room temperature.

Insulin absorption efficacy methods were used to test this formulation on several diabetic patients. Ten diabetic volunteers were asked to fast overnight and not have any breakfast prior to dosing. The patients were challenged with a high calorie meal after the insulin dose. Blood glucose levels were measured for the next 4 hours. Results are shown in Table 1. On day one, patients were given placebo puffs and an oral hypoglycemic agent (Metformin, "Tablets"); on day two, patients were given a 70 unit dose of oral insulin prepared as described above; and on day three, patients were given a 70 unit dose of the present oral insulin formulation. As seen in Table 1, the present oral insulin formulations performed much better than the oral hypoglycemic agents in controlling glucose levels.

TABLE 1 Placebo Tablets Oral-70 Oral-70-2 Repeat Dose Day-I Day 2 Day 3 6.8 6.4 6.6 6.1 6.3 7.8 6.5 7.1 12.2 8.6 8.9 11.3 9.0 9.1 10.7 10.1 8.4 8.2 The procedure was repeated with the following results: TABLE 2 Placebo Tablets Oral-70 Oral-70-2 Repeat Dose Day-1 Day 2 Day 3 6.3 5.9 6.2 6.7 5.4 5.9 6.0 6.7 10.5 8.4 8.4 10.3 8.2 8.4 9.1 6.8 7.2 5.8 5.9 6.9 5.3 5.3 6.4 5.1 5.2 6.1 4.7 4.7 Example 2 An insulin solution was prepared as described in Example 1. To this solution was added 7 mg sodium lauryl sulfate, 7 mg polyoxyethylene ether (10 lauryl) and 7 mg trihydroxy oxo cholanyl glycine and dissolved completely. Seven mg lecithin, solubilized in a water alcohol solution (7 mg/mL) were then added while stirring at high speed, i.e. 2000 rpm.

The solution was stirred for 30 minutes and then stored at 10 0 C. The resulting mixed micellar solution had about 200 units insulin. To this mixture 5 mg phenol, 5 mg m-cresol and 10 mg glycerin were added.

The solution was pipetted (1 mL/vial) into 10 mL capacity glass vials. The vials were then charged with HFA-134a propellant with a Pamasol 2008 automatic gas filling IDapparatus. The amount of propellant was adjusted to 9 mL shot size in order to deliver 2 0 O units insulin per actuation of the aerosol vial. The valves of the vials were designed to cl Sdeliver 100 pL spray per actuation, containing 2 units insulin. The formulation in the glass vial, including the propellant, was in a single phase, i.e. was homogeneous.

c The aerodynamic particle size was determined by an 8-stage USP Anderson® Cascade IDImpactor Mark II. The impactor was cleaned with methanol and air dried at 30'C. Glass Cx, fibre filters were placed on the collection plates. The actuator was attached to the Omouthpiece of the impactor and assembled onto the USP induction port and jet stages. A tcl IN vacuum pump was connected and the air flow rate set to 28.3 liters per minute. The vial Owas primed by shaking for 10 seconds and actuated twice to waste. The shot was delivered by discharging the actuator into the mouthpiece and repeating 25 times. The deposited insulin was collected by rinsing the mouthpiece with 0.6 mL EDTA in 10 mL water at pH 8.7. The filters were removed and placed in scintillation vials and sonicated for 15 minutes. The quantity of insulin was then analyzed using RP-HPLC. The results are shown in Table 3 (2 units per actuation) and 4 (4 units per actuation).

TABLE3 Stage No. 0 1 2 3 Volume (mL) 10 10 10 Mass (mg) 0.79 0.81 0.78 Units 10.4 10.0 10.0 Actuation 5 5 Units per actuation 2.0 2.0 2.1 Particle size (pIm) 8.8 5.8 5.7 not determined/detected TABLE 4 Stage No. 0 1 2 3 Volume (mL) 10 i0 10 Mass (mg) 0.79 0.81 0.78 Units 20.7 21.0 20.1 IActuation 5 5

C

Units per actuation 4.15 4.18 4.01 Particle size (pm) 9 5.8 4.7 not determined en3 c Based on these tests, the average particle size was determined to be about 7 pm, and Va IN stages 3-8, not all of which are shown, revealed no insulin deposition, indicating that omost particles were larger than about 6 pm. This suggests that there would be no deep N lung deposition of the formulation and that most of the formulation would be deposited in

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Va othe buccal cavity.

Further tests were conducted to determine the shot size accuracy, by firing shots into thiel tubes and weighing the tubes before and after the sample collection. The tests showed the shots for 2 units per actuation weighed between 0.075 and 0.083 grams, i.e. within about The tests showed the shots for 4 units per actuation weighed between 0.076 and 0.083 grams, i.e. within about The tests showed the shots for 6 units per actuation weighed between 0.070 and 0.082 grams, i.e. within about HPLC analysis showed the doses delivered to be from 2.01 units to 2.07 units for 2 units per actuation, from 3.9 units to 4.4 units for 4 units per actuation, and from 5.8 units to 6.3 units for 6 units per actuation.

Ten diabetic volunteers were asked to fast overnight and not have any breakfast prior to dosing. On the first day, the volunteers were given 10 units insulin by injection (regular fast acting insulin, obtained from Eli Lilly). On the second day, the volunteers were given units insulin of this example (10 puffs of 6 units each) into the mouth, without inhalation. Plasma insulin levels were measured at intervals by the RIA method for 3 hours. The average results, in micromoles per ml, are shown in Table 5. Blood glucose levels were also monitored at intervals using Bayer's glucometer Elite for 3 hours. The average results, in millimoles per liter, are shown in Table 6.

TABLE Time*: 0 15 30 45 60 90 120 150 180 Injection: 10 9.1 11 16 31 45 32 25 Spray: 8.7 12.1 19.8 28 27 36 29 21 13 time in minutes This test indicated that the direct insulin injection method and the spray method for delivering the present formulations resulted in comparable plasma insulin levels.

TABLE6 Time*: 0 15 30 45 60 90 120 150 180 Injection: 6.1 6.0 5.9 5.5 5.1 4.5 3.8 4.2 4.4 Spray: 6.6 6.3 5.8 5.2 4.8 4.9 4.5 5.0 5.3 time in minutes This test indicated that the direct insulin injection method and the spray method for delivering the present formulations resulted in comparable blood glucose level.

Tests were also conducted with 40 units of spray at 10 puffs of 4 units each, and compared to 10 units injected by measuring plasma levels and glucose levels as above.

The results are shown in Table 7 (plasma) and 8 (glucose).

TABLE 7 Time*: 0 15 30 45 60 90 120 150 180 Injection: 9 9 13 19 34 45 42 35 24 Spray: 10 13 18.5 27 30 33 29 19 14 time in minutes This test indicated that the direct insulin injection method and the spray method for delivering the present formulations resulted in comparable plasma insulin.

TABLE 8 Time*: 0 15 30 60 90 120 150 180 Injection: 5.8 6.0 5.9 5.5 5.0 4.5 4. 1 3.9 Spray: 6.0 5.7 5.4 5.0 5.1 4.7 4.5 4.2 time in minutes IDThis test indicated that the direct insulin injection method and the spray method for

C

O delivering the present formulations resulted in comparable glucose levels.

Cl CtExample 3 en3 C An insulin solution was prepared as described in Example 1. To this solution was added 30.4 mg sodium lauryl sulfate per ml of insulin solution, 30.4 mg polidocanol 9 lauryl Va r- ether per ml of insulin solution and 10.0 mg polylysine per ml of insulin solution, and the Ocompounds dissolved completely. 15.2 mg triolein per ml of insulin solution was then C added while stirring at high speed, i.e. 2000 rpm. The solution was stirred for 30 minutes and then stored at 10C. The resulting solution was a mixed micellar solution. To this C mixture 15.2 mg m-cresol per ml of insulin solution were added.

The solution was pipetted (1 mL) into glass vials. The vials were then charged with 10.8 g HFA 134a propellant per vial, with a Pamasol® 2008 automatic gas filling apparatus. The valves of the vials were designed to deliver 100 gL spray per actuation, containing 6 units insulin. The formulation in the glass vial including the propellant, was in a single phase, i.e. was homogeneous.

Ten diabetic volunteers were asked to fast overnight and not have any breakfast prior to dosing. On the first day, the volunteers were given 10 units insulin by injection. On the second day, the volunteers were given 60 units insulin of this example (10 puffs of 6 units each) into the mouth, without inhalation. Plasma insulin levels were measured at intervals by the RIA method for 3 hours. The average results, in micromoles per ml, are shown in Table 9. Blood glucose levels were also monitored at intervals using Bayer's glucometer Elite for 3 hours. The average results, in millimoles per liter, are shown in Table TABLE9 Time*: 0 15 30 45 60 90 120 150 180 Injection: 9 9.1 14 20 40 48 39 34 27 Spray: 10 15.1 22 32 47 36 27 21 19 time in minutes This test indicated that the direct insulin injection method and the spray method for delivering the present formulations resulted in comparable plasma insulin levels.

TABLE Time*: 0 15 30 45 60 90 120 150 180 Injection: 6.6 6.5 6.1 5.5 4.9 4.5 3.8 3.5 4.4 Spray: 6.8 5.9 5.2 4.8 4.3 3.9 4.5 5.7 5.3 time in minutes This test indicated that the direct insulin injection method and the spray method for delivering the present formulations resulted in comparable glucose levels.

Example 4 An insulin solution was prepared as described in Example 1. The solution was diluted with distilled water until there were 600 units insulin per ml of solution. One ml portions were then transferred to 10 mL capacity glass vials, which were then charged with 10.8 g HFA 134a propellant using a Pamasol® 2008 semi-automatic gas filling apparatus.

The gas phase and the aqueous phase were observed to be distinctly separate. Even shaking of the vials did not appear to homogenize the formulation.

Tests were conducted to determine the shot size accuracy, by firing shots into thiel tubes and weighing the tubes before and after the sample collection. The tests showed five consecutive shots for 6 units per actuation weighed 0.094, 0.110, 0.200, 0.150 and 0.050 grams, i.e. within about ±60% of the average. This compares with in Example 2 (which describes a formulation within the scope of the present invention).

HPLC analysis showed the average doses delivered to be 5.4 units per actuation from shots 5-10, 7.1 units per actuation from shots 45-50 and 8.6 units per actuation from shots 85-90.

These results showed that uniform dose delivery is achievable with the micelle-forming ingredients of the present invention, but not without, based upon the results of this Example as compared with the results of Example 2.

IN Example Ten ml of concentrated insulin containing 10,000 units per ml were placed in a glass c beaker. To this solution was added 7 mg sodium lauryl sulfate, 7 mg polyoxyethylene eC ether (10 lauryl), 7 mg trihydroxy oxocholanyl glycine and 7 mg lecithin. The components were stirred until they were completely dissolved. Seven mg phenol and 7 IN mg m-cresol were added to the solution and mixed thoroughly.

SOne mI portions of the solution were pipetted into 10 mL capacity glass vials. The vials C had metered dose valves thereon. The vials were then charged with HFA 134a propellant Swith Pamasol® 2008 gas filling apparatus. The amount of propellant was adjusted to 9 0 Cl mL per vial in order to deliver 10 units of insulin per actuation of the valve (100 iL shot/actuation). The formulation, in the glass vial, including the propellant, was in a single phase, i.e. was homogeneous.

Ten diabetic human patients fasted overnight and did not have a breakfast prior to dosing.

On the first day, each patient had 7 units regular fast acting insulin, obtained from Eli Lilly, administered by injection. On the second day, each patient was given 70 units of the insulin formulation of this Example (7 puffs of 10 units each) into the mouth, without inhalation. Blood samples were collected and plasma glucose levels were measured at intervals using Bayer's glucometer Elite for 3 hours. The average results, in millimoles per ml, are shown in Table 11. Insulin levels were also monitored at intervals by the RIA method for 3 hours. The average results, in micromoles per liter, are shown in Table 12.

TABLE 11 Time*: 0 15 30 45 60 90 120 150 180 Injection: 6.5 6.3 5.7 5.2 4.8 4.9 3.8 4.5 4.7 Spray: 6.1 6.0 6.0 5.9 5.5 4.5 3.6 4.1 4.4 time in minutes Va c en ci

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cIN

IN

TABLE 12 Time*: 0 15 30 45 60 90 120 150 180 Injection: 8.7 12.1 19.8 29.0 36.0 37.0 33.0 23.0 14.0 Spray: 9.1 11.0 16.0 31.0 45.0 43.0 45.0 32.0 22.0 time in minutes This test indicated that the direct insulin injection method and the spray method of administering the present formulations resulted in comparable insulin levels.

Example 6 Ten ml of concentrated insulin containing 10,000 units per ml were placed in a glass beaker. To this solution was added 15 mg sodium lauryl sulfate, 15 mg chenodeoxycholate, 15 mg trihydroxy oxocholanyl glycine and 7 mg lecithin. The components were stirred until they were completely dissolved. Seven mg phenol and 7 mg m-cresol were added to the solution and mixed thoroughly.

One ml portions of the solution were pipetted into 10 mL capacity glass vials. The vials had metered dose valves thereon. The vials were then charged with HFA 134a propellant with Pamasol® 2008 gas filling apparatus. The amount of propellant was adjusted to 9 mL per vial in order to deliver 10 units of insulin per actuation of the valve (100 PaL shot/actuation). The formulation, in the glass vial, including the propellant, was in a single phase, i.e. was homogeneous.

Ten diabetic patients fasted overnight and did not have a breakfast prior to dosing. On the first day, each patient had 7 units regular fast acting insulin, obtained from Eli Lilly, administered by injection. Fifteen minutes after administering the insulin, each patient was given a 250-calorie Sustacal® drink, which was consumed within 10 minutes. On the second day, each patient was given 70 units insulin of this example (7 puffs of 10 units each) into the mouth, without inhalation. Fifteen minutes after administering the insulin, each patient was given a 250-calorie Sustacal® drink, which was consumed within minutes. Blood samples were collected and plasma glucose levels were measured at intervals, using Bayer's glucometer Elite for 4 hours. The average results, in miIlimoles per ml, are shown in Table 13.

TABLE 13 Time*: 0 15 30 60 90 120 150 180 Injection: 9.2 9.0 9.5 12.3 12.4 12.6 11.3 9.7 Spray: 8.8 8.8 8.7 10.4 12.0 12.4 11.9 10.5 time in minutes These tests indicated that the direct insulin injection method and the spray method for administering the present formulations resulted in comparable blood glucose levels.

Whereas particular embodiments of this invention have been described above for purposes of illustration, it will be evident to those skilled in the art that numerous variations of the details of the present invention may be made without departing from the invention as defined in the appended claims.

Claims (27)

1. A pharmaceutical formulation comprising: an effective amount of a macromolecular pharmaceutical agent in micellar form; an alkali metal alkyl sulfate; r at least three micelle-forming compounds selected from the group comprising lecithin, 0 hyaluronic acid, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic V C acid, linoleic acid, linolenic acid, monoolein, monooleates, monolaurates, borage oil, o evening primrose oil, menthol, trihydroxy oxo cholanyl glycine, glycerin, polyglycerin, C lysine, polylysine, triolein, polyoxyethylene ethers, polidocanol alkyl ethers, chenodeoxycholate, deoxycholate, pharmaceutically acceptable salts thereof, analogues thereof, and mixtures or combinations thereof; and a suitable solvent; wherein the alkali metal alkyl sulfate and each of the micelle-forming compounds are each present in a concentration of between about 0.1 and 20 wt./wt. of the total formulation, and the total concentration of the alkali metal alkyl sulfate and micelle- forming compounds together is less than 50 wt./wt. of the total formulation.

2. A formulation according to claim 1, wherein the alkali metal alkyl sulfate is in a concentration of less than about 5 wt./wt. of the total formulation.

3. A formulation according to claim I or 2, wherein each of said three or more micelle-forming compounds is present in a concentration of between about 0.1 and wt./wt. of the total formulation.

4. A formulation according to any one of claims 1 to 3, wherein the total concentration of the alkali metal alkyl sulfate and micelle-forming compounds together is less than 4 wt./wt. of the total formulation.

A formulation according to any one of claims 1 to 4, wherein the alkali metal alkyl sulfate is an alkali metal C8 to C22 alkyl sulfate. 0

6. A formulation according to claim 5, wherein the alkali metal C8 to C22 alkyl O 0 sulfate is sodium lauryl sulfate.

7. A formulation according to any one of claims 1 to 6, wherein the lecithin is either eC saturated or unsaturated and is selected from the group comprising phosphatidylcholine, phosphatidylserine, sphingomyelin, phosphatidylethanolamine, cephalin, and lysolecithin.

8. A formulation according to any one of claims 1 to 7, wherein one of the micelle- O forming compounds is selected from the group comprising hyaluronic acid, 0 C pharmaceutically acceptable salts of hyaluronic acid, polidocanol alkyl ethers, trihydroxy O oxo cholanyl glycine and pharmaceutically acceptable salts thereof, polyoxyethylene 0 CI ethers and analogues thereof, chenodeoxycholate, and mixtures thereof.

9. A formulation according to claim 8, wherein said salt of hyaluronic acid is selected from the group comprising alkali metal hyaluronates, alkaline earth hyaluronates, and aluminum hyaluronates.

A formulation according to any one of claims 1 to 7, wherein the three micelle- forming compounds are a combination selected from the group comprising: trihydroxy oxo cholanyl glycine, polyoxyethylene ether and lecithin; trihydroxy oxo cholanyl glycine, deoxycholate and glycerin; polidocanol 9 lauryl ether, polylysine and triolein; trihydroxy oxo cholanyl glycine, chenodeoxycholate and glycerin; trihydroxy oxo cholanyl glycine, polyoxyethylene ether and glycerin; polidocanol 10 lauryl ether, sodium glycocholate and lecithin; polidocanol 10 lauryl ether, sodium hyaluronate and lecithin; polidocanol 20 lauryl ether, evening primrose oil and lecithin; and polidocanol 10 lauryl ether, phosphatidyl choline and oleic acid.

11. A formulation according to any one of claims 1 to 10, wherein the pharmaceutical agent is selected from the group comprising insulin, heparin, low molecular weight heparin, hirulog, hirugen, huridin, interferons, cytokines, mono and polyclonal antibodies, immunoglobins, chemotherapeutic agents, vaccines, glycoproteins, bacterial toxoids, hormones, calcitonins, glucagon like peptides, antibiotics, protein based thrombolytic N0 compounds, platelet inhibitors, DNA, RNA, gene therapeutics, antisense oligonucleotides, O O opioids, narcotics, hypnotics, steroids and pain killers.

12. A formulation according to claim 11, wherein the pharmaceutical agent is insulin. Cl

13. A formulation according to any one of claims 1 to 12, wherein the pH of said formulation is between 5 and 8. Va r- Cl

14. A formulation according to any one of claims 1 to 13, wherein the size of said 0 O micelles is greater than 6 pm.

C< A formulation according to any one of claims 1 to 14, wherein said solvent is selected from the group comprising water and ethanol.

16. A formulation according to claim 15, wherein said solvent is water.

17. A mixed micellar pharmaceutical formulation comprising a macromolecular pharmaceutical agent encapsulated in micelles formed with an alkali metal alkyl sulfate and at least three compounds selected from the group comprising lecithin, hyaluronic acid, glycolic acid, lactic acid, chamomile extract, cucumber extract, oleic acid, linoleic acid, linolenic acid, monoolein, monooleates, monolaurates, borage oil, evening primrose oil, menthol, trihydroxy oxo cholanyl glycine, glycerin, polyglycerin, lysine, polylysine, triolein, polyoxyethylene ethers, polidocanol alkyl ethers, chenodeoxycholate, deoxycholate, pharmaceutically acceptable salts thereof, analogues thereof, and mixtures or combinations thereof.

18. A formulation according to any one of claims 1 to 17 further comprising a phenolic compound selected from the group comprising phenol and methyl phenol in a concentration of from 1 to 10 wt./wt. of the total formulation.

19. A formulation according to any one of claims 1 to 18 further comprising a propellant selected from the group consisting of C1-C2 dialkyl ether, butanes, fluorocarbon propellant, hydrogen-containing fluorocarbon propellant, chlorofluorocarbon propellant, hydrogen-containing chlorofluorocarbon propellant, and mixtures thereof. IO

20. A formulation according to claim 19 wherein the propellant is selected from the O 0 group consisting of tetrafluoroethane, tetrafluoropropane, dimethylfluoropropane, g heptafluoropropane, dimethyl ether, n-butane and isobutane. Cn

21. A formulation according to claim 19 or 20 wherein the ratio of pharmaceutical agent to propellant is from 5:95 to 25:75. \0

22. A metered dose device containing a formulation according to any one of claims 1 0 to 21. O I

23. A method of treating a patient comprising administering to said patient an 0 0 effective amount of a pharmaceutical formulation according to any one of claims 1 to 21.

24. A method according to claim 23, wherein said pharmaceutical formulation is sprayed into an oral cavity of said patient using a metered dose device.

A pharmaceutical formulation substantially as hereinbefore described with reference to the Examples.

26. A method of preparing a pharmaceutical formulation substantially as hereinbefore described with reference to the Examples.

27. A method of treating a patient, substantially as hereinbefore described with reference to the Examples. DATED THIS TWENTY-THIRD DAY OF JANUARY 2006 GENEREX PHARMACEUTICALS INC. BY PIZZEYS PATENT AND TRADE MARK ATTORNEYS