Ultra-long acting pharmaceutical composition containing insulin

Abstract

The present invention discloses a stable ultra-long acting injectable pharmaceutical composition comprising insulin glargine, collagen, one or more pharma- ceutically acceptable excipients or carriers at a pH of 2 to 5. The present invention also relates to a process for preparing said composition and its use for the treatment of metabolic disorders, including diabetes.

Description

[Technical Field]

[0001] The present invention relates broadly to the field of biopharmaceuticals, and in particular to ultra-long acting insulin glargine dosage forms. Specifically, the present invention relates to a stabilized injectable solution comprising insulin glargine, collagen, and one or more pharmaceutically acceptable excipients or carriers thereof. The present invention also relates to a method for preparing the composition and its use for the treatment of metabolic disorders, including diabetes mellitus. [Background technology]

[0002] Diabetes is a serious medical condition characterized by the body's inability or insufficient glucose metabolism. In 2019, approximately 9.3% of the global adult population was diagnosed with diabetes, and by 2045, this number is expected to increase to almost 11% (https: / / www.statista.com / statistics / 271464 / percentage-of-diabetics worldwide / ).

[0003] Diabetes mellitus is a leading cause of death worldwide. There are two major types of diabetes: type 1 diabetes, in which damage to pancreatic beta cells leads to insufficient insulin secretion and requires frequent administration of exogenous insulin to sustain life; and It is caused by a lack of endogenous insulin to regulate blood sugar and can be managed through diet, exercise, medication, or insulin injections, although about 20% of cases are classified as type 2 diabetes, which is controlled with insulin injections.

[0004] In both types of diabetes, hypoglycemia frequently occurs due to the use of insulin, because it is very difficult to predict the amount of endogenous insulin normally secreted by the body to regulate blood sugar. Diabetic patients must periodically administer insulin to maintain blood sugar levels within the normal range.

[0005] Insulin injections are prescribed to patients with diabetes. Insulin is a natural hormone secreted to regulate blood glucose levels. In healthy individuals, insulin is released into the blood by the pancreas when blood glucose levels rise. Increased blood glucose levels after a meal are quickly regulated by increased insulin secretion. Insulin plays an important role in converting excess blood glucose into glycogen, which is then stored in the liver.

[0006] Since the introduction of insulin in the 1920s, there have been sustained efforts to improve the treatment of diabetes. To avoid extreme blood sugar levels, diabetics receive multiple injections of insulin, often with meals.

[0007] Insulin is a polypeptide consisting of 51 amino acids, divided into two amino acid chains: chain A with 21 amino acids and chain B with 30 amino acids. The two chains are linked to each other through two disulfide bridges. Insulin preparations have long been used to treat diabetes.

[0008] Traditionally, rapid-acting generic insulin preparations or its intermediate-acting protamine insulin preparations were used to treat diabetic patients. Over time, new insulin analogs and derivatives have been developed. These insulin analogs and derivatives have one or more amino acid positions changed in the human insulin structure or have different amino acid chain lengths.

[0009] Many insulins, insulin analogues and derivatives are commercially available. Commonly used insulins, insulin analogues or insulin derivatives can be classified as follows:

[0010] Rapid-acting insulin analogs (Bolus): e.g., insulin aspart (Novolog®); insulin lispro (Humalog®); insulin glulisine (Aprida®), rapid-acting human insulin (Viaject®). Such analogs have an onset of action within 5 to 15 minutes of administration and are active for 3 to 4 hours.

[0011] Short-acting insulin (Bolus): For example, regular insulin (Humulin® or Novolin®). Regular insulin begins to act within 30 minutes of administration and has a duration of action of about 5 to 8 hours.

[0012] Intermediate-acting insulins, such as isophane insulin, aspartoprotamine, and lisproprotamine, act 1 to 3 hours after administration, with a duration of action of 16 to 24 hours.

[0013] Long-acting insulins (Basal): e.g., insulin glargine, insulin degludec, and insulin detemir. Such analogs begin to act within 1 to 2 hours, and their duration of action varies from about 12 to about 24 hours.

[0014] Premixed insulins: for example, a mixture of NPH and regular insulin. There are various formulations with different mixing ratios of premixed insulins. The action of such premixed preparations begins within about 30 minutes. Premixed insulins contain insulins of the same type. Two other types of insulin cannot be mixed. That is, insulin lispro cannot be mixed with insulin detemir, insulin aspart, or insulin glargine. Premixed insulin lispro dosage forms can only mix two forms of insulin lispro: insulin lispro regular and insulin lispro protamine.

[0015] Insulin glargine is a long-acting basal insulin administered once daily. It is formulated in an acidic solvent and has an altered isoelectric point. It is isoelectric at physiological pH and precipitates after subcutaneous injection, forming an amorphous depot. The gradual redissolution of this subcutaneously formed depot is the primary delay mechanism. The long duration of action of insulin glargine (up to 24 hours) is directly related to the formation of fine precipitate particles; the dissociation rate of the fine precipitate from the insulin glargine monomer units; and the slow absorption rate of insulin glargine from the subcutaneous tissue. However, in many patients, insulin glargine does not persist over 24 hours. The drug precipitates after injection into the subcutaneous tissue and is gradually redissolved and absorbed. Although it has a "peak-less" action, the drug is associated with interindividual variability, and many patients experience peak action, especially at high doses.

[0016] Another disadvantage of insulin glargine is that, unlike isophane insulin, it cannot be mixed with soluble insulin, resulting in precipitation. Therefore, many patients using dual (mixed) insulins must increase the number of daily injections or switch to a basal-bolus injection regimen. Furthermore, many patients develop insulin resistance after prolonged use of long-acting insulin, necessitating increased insulin injection doses. In vitro studies have shown that glargine preferentially binds to insulin-like growth factor-1 (IGF-1) receptors over insulin receptors. This may indicate the potential for mitogenic stimulation with prolonged use. Therefore, there is a need for insulin glargine formulations that not only exhibit a longer duration of action but also reduced interindividual variability and a less mitogenic effect.

[0017] Biomaterials are widely utilized in the drug and pharmaceutical fields for efficient drug delivery in the body. Among known biopolymers, collagen has emerged as one of the most attractive choices due to its excellent biocompatibility and biodegradability, weak antigenicity, and known "molecular structure, biological properties, and mode of interaction with the body" (Friess, 1998; Lee et al., 2001).

[0018] Collagen, the most abundant protein in mammals, is the major structural protein, accounting for approximately 30% of the total body protein in all vertebrates. Collagen makes up more than 90% of the extracellular proteins in muscle and bone, and more than 50% of the extracellular proteins in skin. The primary sources of collagen today are the skin of animals such as cattle or pigs, primarily the Achilles tendons of cattle or horses.

[0019] To date, 28 types of collagen have been identified and described, the most common types being: Type 1: Main component of bone tissue Type 2: Main component of cartilage Type 3: Main component of reticular fibers Type 4: The main component of the lamina densa, the epithelial secretory layer of the basement membrane Type 5: Found primarily on cell surfaces, hair, and the placenta.

[0020] There is a large amount of prior art that discloses pharmaceutical compositions containing the binding affinity and combination of insulin and collagen.

[0021] U.S. Patent Application No. 20130225492 discloses a pharmaceutical composition containing a drug and collagen, which is easy to handle and has sustained release properties. The sustained release pharmaceutical composition contains a drug, collagen, and at least one sugar selected from monosaccharides, disaccharides, trisaccharides, and tetrasaccharides.

[0022] U.S. Patent No. 5,922,356 B2 discloses that a sustained-release preparation containing a therapeutically active substance as an active ingredient, collagen as a drug carrier, and glycosaminoglycan as an additive is used for the treatment or prevention of disease.

[0023] Japanese Patent No. 0543453 discloses a topical sustained-release preparation for promoting wound healing, which is obtained by mixing a carrier containing collagen as an essential component with a physiologically active substance having wound healing promoting activity.

[0024] Japanese Patent No. 56122317 discloses that when the concentration of collagen decreases, the decomposition of a drug containing gelled collagen is accelerated and the release rate of the drug becomes faster.

[0025] Yaoi et al. (1991) reported on the binding efficiency of insulin to collagen by analyzing the ability of eight extracellular matrix proteins, namely fibronectin, vitronectin, laminin, and collagen types 1, 2, 3, 4, and 5, to bind to insulin.

[0026] Manolache et al., 2016, discloses a gel complex and matrix comprising collagen gel, zinc oxide, and insulin glargine at pH 7.4.

[0027] U.S. Patent No. 6,468,959 discloses a dry pellet as a dosage form that can be administered orally, bucally, sublingually, or nasally, containing at least one peptide drug having a given net charge dispersed in a matrix containing a component selected from the group consisting of gelatin, fractionated gelatin, collagen hydrolysate, cross-linked gelatin, and mixtures thereof, wherein the hydrophilic molecular group components possess a net charge opposite enough to form a pseudocoacervate with the peptide drug.

[0028] U.S. Patent Application Publication No. 20140213963 discloses a biocompatible insulin delivery device comprising an insulin reservoir and a glucose-responsive plug sealing the reservoir, optionally covering the exposed surface of the glucose-responsive plug and a protective microporous membrane additionally sealing the reservoir, wherein the plug comprises a polymer matrix having a stimulus-responsive component and an inorganic component adapted to alter the porosity of the plug in response to a stimulus, and wherein the plug functions to release insulin from the reservoir in response to hyperglycemic concentrations and to prevent insulin from being released from the reservoir in response to hypoglycemic glucose concentrations.

[0029] Compositions containing insulin and collagen are known in the prior art. These compositions are in the form of gels, pellets, or implants. The administration of insulin as a gel or implant requires specialized syringes or devices, reducing ease of administration. Ease of administration is important because diabetic patients must administer insulin repeatedly. Administration of gels or implants induces pain at the injection site, frequently causes swelling, and is inconvenient for patients, resulting in low compliance. Furthermore, specialized syringes or devices for administering such medications increase the overall cost of the product and are not suitable for many patients. Oral administration of insulin as pellets results in high variability in absorption rates. Oral administration of insulin often results in unpredictable pharmacokinetic responses. Summary of the Invention [Problem to be solved by the invention]

[0030] Therefore, there is a need for insulin glargine and collagen dosage forms that are not only easy to administer using standard pens and syringes, but also have increased bioavailability, a longer duration of action, and reduced immunogenicity compared to commercially available insulin glargine compositions of the same dose.

[0031] None of the prior art discloses an injectable dosage form comprising insulin glargine, collagen and one or more pharmaceutically acceptable excipients having a pH of 2 to 5, which has a longer duration of action and greater bioavailability than commercially available insulin glargine formulations at the same strength. [Means for solving the problem]

[0032] The term "insulin" as used herein includes mammalian insulin, insulin analogs or derivatives.

[0033] The term "insulin analogue or derivative" as used herein includes analogues or derivatives of the naturally occurring insulin molecule, i.e., insulin analogues or derivatives that have been modified from the structure of human or animal insulin and have altered physicochemical, pharmacokinetic, and pharmacodynamic properties.

[0034] One aspect of the present invention provides a stable, ultra-long-acting injectable pharmaceutical composition comprising an insulin analog or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof, collagen, and one or more pharmaceutically acceptable excipients or carriers. The composition has a pH of 2 to 5, is a solution upon administration, and forms a depot at physiological pH immediately after administration. The composition exhibits increased bioavailability and a longer duration of action compared to commercially available injectable compositions containing "insulin analogs or derivatives having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof" at the same dosage.

[0035] One aspect of the present invention provides a stable, ultra-long acting injectable pharmaceutical composition comprising insulin glargine or a physiologically acceptable salt thereof; collagen; and one or more pharmaceutically acceptable excipients or carriers, wherein the pharmaceutical composition has a pH of 2 to 5; the pharmaceutical composition is in solution upon administration and forms a depot at physiological pH immediately after administration; and the pharmaceutical composition exhibits increased bioavailability and a longer duration of action compared to commercially available injectable pharmaceutical compositions containing insulin glargine at the same concentration or dose.

[0036] One aspect of the present invention provides a stable, ultra-long acting injectable pharmaceutical composition comprising insulin glargine or a physiologically acceptable salt thereof; collagen; and one or more pharmaceutically acceptable excipients or carriers, wherein the pharmaceutical composition has a pH of 2 to 5; the pharmaceutical composition is in solution upon administration and forms a depot at physiological pH immediately after administration; and the pharmaceutical composition exhibits increased bioavailability and a duration of action of 24 hours or more.

[0037] One aspect of the present invention provides a stable, ultra-long acting injectable pharmaceutical composition comprising insulin glargine or a physiologically acceptable salt thereof; human collagen; and one or more pharmaceutically acceptable excipients or carriers, wherein the pharmaceutical composition has a pH of 2 to 5; the pharmaceutical composition is in solution upon administration and forms a depot at physiological pH immediately after administration; and the pharmaceutical composition exhibits increased bioavailability and a longer duration of action compared to commercially available injectable compositions containing insulin glargine at the same concentration or dose.

[0038] One aspect of the present invention provides a stable, ultra-long-acting injectable pharmaceutical composition comprising an insulin analog or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof; collagen; and one or more pharmaceutically acceptable excipients or carriers, wherein the pharmaceutical composition has a pH of 2 to 5; the pharmaceutical composition is in solution upon administration and forms a depot at physiological pH immediately after administration; the pharmaceutical composition exhibits increased bioavailability and a longer duration of action compared to commercially available injectable pharmaceutical compositions containing the "insulin analog or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof" at the same dose; and the pharmaceutically acceptable excipients or carriers are selected from the group consisting of a stabilizer, an isotonic agent, a surfactant, a buffer, a preservative, an antioxidant, an acidifier, a pH adjuster, an organic solvent, an aqueous solvent, a solubilizer, and combinations thereof.

[0039] One aspect of the present invention provides a method for treating type 1 and type 2 diabetes in a patient, comprising administering a stable, ultra-long-acting injectable pharmaceutical composition comprising an insulin analog or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof, collagen, and one or more pharmaceutically acceptable excipients or carriers, wherein the pharmaceutical composition has a pH of 2 to 5; the pharmaceutical composition is in solution upon administration and forms a depot at physiological pH immediately after administration; the pharmaceutical composition exhibits increased bioavailability and a longer duration of action compared to an injectable pharmaceutical composition containing the insulin analog or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof, at the same dose; and the pharmaceutical composition is administered two to three times per week.

[0040] One aspect of the present invention provides a method for preparing a stable, ultra-long acting injectable solution comprising an insulin analog or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof; collagen; and one or more pharmaceutically acceptable excipients or carriers having a pH of 2 to 5, the method comprising the steps of: a) dissolving an accurately weighed amount of zinc-containing insulin glargine crystals in water for injection with the aid of a few μL of 1 M HCl; b) preparing a buffer of pH 2 to 5; c) dissolving an isotonicity agent in a buffer; d) diluting the concentrated insulin glargine of step (A) with the buffer of step b); e) adding collagen to the solution of step d) under constant stirring; f) adjusting the final pH of the solution to between 2 and 5. This specification includes the disclosures of Appendices 1 to 14 below. (Appendix 1) A stable, ultra-long-acting injectable pharmaceutical composition comprising an insulin analog or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof; collagen; and one or more pharmaceutically acceptable excipients or carriers, The pharmaceutical composition has a pH of 2 to 5; The pharmaceutical composition is in solution upon administration and forms a depot at physiological pH immediately after administration; The pharmaceutical composition is a stable, ultra-long-acting injectable pharmaceutical composition that exhibits increased bioavailability and longer duration of action compared to an injectable pharmaceutical composition containing the "insulin analogue or derivative having an isoelectric point of 5 to 8.5" at the same dose. (Appendix 2) 2. The stable, ultra-long-acting injectable pharmaceutical composition according to claim 1, wherein the "insulin analogue or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof" is insulin glargine. (Appendix 3) 3. The stable, ultra-long-acting injectable pharmaceutical composition of claim 2, wherein the insulin glargine is present in a concentration range of 100 IU to 1,000 IU / ml. (Appendix 4) 2. The stable, ultra-long-acting injectable pharmaceutical composition according to claim 1, wherein the collagen is human collagen. (Appendix 5) The stable, ultra-long-acting injectable pharmaceutical composition described in Appendix 1, characterized in that the pharmaceutical composition has a duration of action of 24 hours or more. (Appendix 6) 2. The stable, ultra-long-acting injectable pharmaceutical composition of claim 1, wherein the pharmaceutical composition is administered two or three times a week. (Appendix 7) The stable, ultra-long-acting injectable pharmaceutical composition described in Appendix 1 is characterized in that the pharmaceutical composition has lower immunogenicity than a commercially available injectable pharmaceutical composition containing the same dose of the "insulin analogue or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof." (Appendix 8) The stable ultra-long-acting injectable pharmaceutical composition described in Appendix 1, characterized in that the pharmaceutically acceptable excipient is selected from the group consisting of stabilizers, isotonicity agents, surfactants, buffers, preservatives, antioxidants, acidifying agents, pH adjusters, organic solvents, water-soluble solvents, solubilizers, and combinations thereof. (Appendix 9) A stable, ultra-long-acting injectable pharmaceutical composition comprising an insulin analog or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof; collagen; and one or more pharmaceutically acceptable excipients or carriers, The pharmaceutical composition has a pH of 2 to 5; The pharmaceutical composition is in solution upon administration and forms a depot at physiological pH immediately after administration; The pharmaceutical composition is a stable, ultra-long-acting injectable pharmaceutical composition characterized by exhibiting increased bioavailability and a longer duration of action compared to commercially available injectable pharmaceutical compositions containing the "insulin analogue or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof" at the same dose for use in treating one or more metabolic disorders. (Appendix 10) 10. The stable, ultra-long acting injectable pharmaceutical composition according to claim 9, wherein the metabolic disorder is selected from the group consisting of insulin resistance syndrome, diabetes, hyperlipidemia, fatty liver disease, cachexia, obesity, atherosclerosis, and arteriosclerosis. (Appendix 11) 10. A method for treating type 1 or type 2 diabetes in a patient in need thereof, comprising administering to the patient the pharmaceutical composition of claim 1. (Appendix 12) 1. A method for preparing a stable, ultra-long acting injectable solution comprising an insulin analog or derivative having an isoelectric point of 5 to 8.5, a physiologically acceptable salt, collagen, and one or more pharmaceutically acceptable excipients or carriers having a pH of 2 to 5, the method comprising: a) solubilizing insulin glargine with hydrochloric acid (HCl) solution and water for injection; b) preparing a buffer of pH 2 to 5; c) dissolving an isotonicity agent in the buffer of step b); d) diluting the insulin glargine solution of step a) with the buffer of step c); e) adding collagen to the solution of step d) under constant stirring; and f) adjusting the final pH of the solution to 2 to 5. (Appendix 13) 13. A method for producing a stable, ultra-long-acting injectable solution according to claim 12, wherein the "insulin analogue or derivative having an isoelectric point of 5 to 8.5" is insulin glargine. (Appendix 14) 1. A method for modulating the release of an insulin analog or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof, from a pharmaceutical composition, comprising: The method includes the steps of formulating a pharmaceutical composition at a pH of 2 to 5, the pharmaceutical composition comprising an insulin analog or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof; collagen; and one or more pharmaceutically acceptable excipients or carriers having a pH of 2 to 5, and precipitating the composition at physiological pH immediately after administration. [Effects of the Invention]

[0041] The present invention discloses a stable, ultra-long acting injectable solution comprising insulin glargine, collagen and one or more of these excipients at a pH of 2 to 5. The present invention also discloses a method for preparing said composition and its use for the treatment of metabolic disorders, including diabetes. [Brief explanation of the drawings]

[0042] [Figure 1] 1 shows a standard curve of insulin glargine in the presence of collagen. [Figure 2] 1 shows a simulation of plasma insulin concentrations following once-daily administration of Gla-100 or Gla-300 according to the prior art. (Reference: Modeling of Subcutaneous Absorption of Long-Acting Insulin Glargine in Type 1 Diabetes, Michele et al.: IEE Trans Biomed Eng 202;67(2):624-631.) [Figure 3] 1 shows an "in vitro model of insulin glargine release" used in the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0043] While investigating other dosage forms to produce a pharmaceutical composition containing insulin glargine that not only exhibits a duration of action of 24 hours or more, but also is easy to administer, has higher bioavailability, and is less immunogenic, the inventors observed that when human type I collagen was mixed with insulin glargine at an acidic pH, the formation of M-1 and M-2, soluble metabolites of insulin glargine, at physiological pH was significantly reduced (M-1 and M-2 are insulin metabolites generated when insulin glargine is exposed to proteases in saline and treated to resemble the composition of interstitial fluid). Because these metabolites of insulin glargine are responsible for hypoglycemic activity, any drug dosage form that can reduce the formation of these metabolites has a direct correlation with the degree of sustained drug release and dissolution flatness. The formation of a pharmaceutical depot at the injection site based on the in vitro kinetics of M-1 and M-2 formation is interpreted as extending the half-life of the circulating active drug and improving its pharmacological efficacy.

[0044] The composition of the present invention is acidic and forms a depot at physiological pH after administration in vivo. The inventors observed that human type I collagen molecules, a triple-helical protein, trap insulin at the injection site (physiological pH) and form an additional barrier for already crystallized insulin glargine at physiological pH 7.4 to inhibit the action of proteases in the interstitial fluid within the extracellular matrix at the injection site. A two-compartment model was proposed for insulin glargine, which is absorbed through the systemic blood circulation. The first compartment confirms the redissolution of precipitates, the formation of hexamers, and the subsequent conversion of the hexamers to monomers. The second compartment confirms the transport of insulin to the blood via vascular capillaries in the subcutaneous tissue. This confirmed that insulin release is controlled by the additional barrier of collagen at physiological pH 7.4. Furthermore, it was confirmed that collagen inhibits proteases that degrade insulin at the injection site. This protease inhibition increases the bioavailability of insulin in the blood, meaning that collagen can cooperate with insulin to promote blood glucose regulation. Therefore, the pharmaceutical composition of the present invention has superior bioavailability compared to commercially available compositions containing insulin glargine at the same concentration.

[0045] The pharmaceutical composition of the present invention is a solution because both insulin glargine and collagen are soluble at acidic pH. The solution of the present invention is easy to administer using a standard syringe or pen and does not require any special syringe or device for administration. The easy administration by injection increases patient compliance. In addition, the pharmaceutical composition of the present invention can be easily administered using existing pens and syringes, making it cost-effective and not placing an additional burden on patients.

[0046] As used herein, the term "pharmaceutical composition" means a mixture containing therapeutic compounds that is administered to a mammal, e.g., a human, to prevent, treat, or control a particular disease or condition affecting the mammal.

[0047] As used herein, "collagen" refers to the fibrous protein that makes up bone, cartilage, muscle, and other connective tissues. As used herein, "collagen" includes, but is not limited to, collagen types 1, 2, 3, 4, and 5, human collagen, and processed or modified collagen.

[0048] As used herein, the term "buffer" refers to a solution containing a weak acid and its salt or a weak base and its salt, which is resistant to changes in pH. As used herein, "buffer" includes, but is not limited to, phosphate, acetate, citrate, arginine, glycylglycine, or Tris (i.e., 2-amino-2-hydroxymethyl-1,3-propanediol) buffers and their corresponding salts, and combinations thereof.

[0049] As used herein, "preservative" refers to a compound that can be used to prevent the growth of fungi and other microorganisms. As used herein, "preservative" includes, but is not limited to, benzoic acid, butylparaben, ethylparaben, methylparaben, propylparaben, sodium benzoate, sodium propionate, benzalkonium chloride, benzethonium chloride, benzyl alcohol, cetylpyridinium chloride, chlorobutanol, phenol, phenylethyl alcohol, 2-phenoxyethanol, phenylmercuric nitrate, thimerosal, metacresol, and combinations thereof.

[0050] As used herein, "tonicity adjusting agent" refers to a physiologically acceptable compound that prevents water circulation across tissue cell membranes in contact with the formulation, imparting a suitable tonicity to the formulation. "Tonicity adjusting agents" include compounds such as glycerin, which are commonly used for such purposes in known concentrations. Other usable tonicity adjusting agents include salts, e.g., sodium chloride, dextrose, lactose, and combinations thereof.

[0051] As used herein, "pH adjuster" refers to a combination of an acid and an alkali. The "pH adjuster" as used herein may be selected from the group consisting of o-phosphoric acid, citric acid, acetic acid, succinic acid, lactic acid, gluconic acid, tartaric acid, 1,2,3,4-butanetetracarboxylic acid, fumaric acid, or malic acid. The alkali may be selected from the group consisting of sodium hydroxide, potassium hydroxide, sodium hydroxide, ammonium hydroxide, magnesium oxide, calcium hydroxide, calcium carbonate, magnesium carbonate, magnesium aluminum silicate, diethanolamine, monoethanolamine, sodium carbonate, sodium bicarbonate, or triethanolamine, and combinations thereof.

[0052] As used herein, "solubilizing agent" refers to a substance that can solubilize or partially solubilize a therapeutic compound and / or polymer. Suitable "solubilizing agents" as used herein include, but are not limited to, humectants such as polysorbates and poloxamers, nonionic and ionic surfactants, food acids and bases (e.g., sodium bicarbonate), polyhydric alcohols, alcohols, and combinations thereof.

[0053] As used herein, "acidifying agent" refers to a compound that donates protons or hydrogen ions and / or accepts electrons. Suitable acidifying agents include, but are not limited to, formic acid, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, tartaric acid, diatrizoic acid, glutamic acid, lactic acid, maleic acid, succinic acid, acetic acid, citric acid or anhydrous citric acid, and combinations thereof, including in the form of particulate solids.

[0054] As used herein, suitable "organic solvents" are selected from the group consisting of, but not limited to, N-methylpyrrolidone (NMP), dichloromethane, dimethylformamide (DMF), dimethylacetamide (DMAC), acetonitrile, tetrahydrofuran, dioxane, methanol, ethanol, isopropanol, tert-butanol, and combinations thereof.

[0055] Suitable "water-soluble solvents" as used herein include water, water for injection, and water and alcohol mixtures.

[0056] As used herein, "surfactants" refers to fatty acid esters or ethers with polyhydric alcohols such as glycerol, sorbitol, etc. (Span®, Tween®, in particular Tween® 20 and Tween® 80, Myrj®, Brij®, Cremophore® or poloxamers, Pluronics® and Tetronics®, polysorbates (Tween TM ), sodium dodecyl sulfate (sodium lauryl sulfate), lauryldimethylamine oxide, cetyltrimethylammonium bromide (CTAB), polyethoxylated alcohol polyoxyethylene sorbitan, octoxynol (Triton X100 TM ), N,N-dimethyldodecylamine-N-oxide, hexadecyltrimethylammonium bromide (HTAB), polyoxyl 10 lauryl ether, Brij 721 TM , bile salts (sodium deoxycholate, sodium cholate), polyoxyl castor oil (Cremophor TM ), Nonylphenol ethoxylate (Tergitol TM ), cyclodextrin, lecithin, methylbenzethonium chloride (Hyamine TM ) and combinations thereof.

[0057] As used herein, an "antioxidant" may be selected from the group consisting of ascorbate (sodium / acid), sodium bisulfite, butylated hydroxyanisole (bha), butylated hydroxytoluene (bht), cysteine / cysteine ​​hydrochloride, sodium dithionite (sodium hydrosulfite, sodium sulfoxylate), gentisic acid, gentisic acid ethanolamine, sodium glutamate, glutathione, sodium formaldehyde sulfoxylate, potassium metabisulfite, sodium metabisulfite, methionine, monothioglycerol (thioglycerol), propyl gallate, sodium sulfite, α-tocopherol, α-tocopherol hydrogen succinate, sodium thioglycolate, and combinations thereof.

[0058] As used herein, "one or more stabilizers" includes, but is not limited to, surfactants, antioxidants, preservatives, solubilizers, esterase inhibitors, and combinations thereof. The one or more stabilizers may be fatty acid esters and ethers with polyhydric alcohols such as glycerol, sorbitol (Span®, Tween®, especially Tween® 20 and Tween® 80, Myrj®, Brij®, Cremophore®, or poloxamers, Pluronics® and Tetronics®, polysorbates (Tween TM ), sodium dodecyl sulfate (sodium lauryl sulfate), lauryldimethylamine oxide, cetyltrimethylammonium bromide (CTAB), glycerol, polyethoxylated alcohol polyoxyethylene sorbitan, octoxynol (Triton X100 TM ), N,N-dimethyldodecylamine-N-oxide, hexadecyltrimethylammonium bromide (HTAB), polyoxyl 10 lauryl ether, Brij 721 TM , bile salts (sodium deoxycholate, sodium cholate), polyoxyl castor oil (Cremophor TM ), Nonylphenol ethoxylate (Tergitol TM), cyclodextrin, lecithin, methylbenzethonium chloride (Hyamine TM ), Benzoic Acid, Zinc as Zinc Oxide, Butylparaben, Ethylparaben, Methylparaben, Propylparaben, Sodium Benzoate, Sodium Propionate, Benzalkonium Chloride, Polyvinyl Alcohol, Benzethonium Chloride, Benzyl Alcohol, Cetylpyridinium Hydrochloride, Chlorobutanol, Phenol, Phenylethyl Alcohol, 2-Phenoxyethanol, Phenylmercuric Nitrate, Thimerosal, Meta-Cresol, Ascorbate (Sodium / Acid), Sodium Bisulfite, Butylated Hydroxyanisole (BHA), Butylated Hydroxytoluene (BHT), Cysteine / Cysteine ​​Sulfate HCl, Dithionite Na sodium (sodium hydrosulfite, sodium sulfoxylate), gentisic acid, gentisic acid ethanolamine, sodium glutamate, glutathione, sodium formaldehyde sulfoxylate, potassium metabisulfite, sodium metabisulfite, methionine, monothioglycerol (thioglycerol), propyl gallate, sodium sulfite, α-tocopherol, α-tocopherol hydrogen succinate, sodium thioglycolate, esterase inhibitors such as pancreatic secretion inhibitors, protease inhibitors, and serine esterase inhibitors such as aprotinin, and combinations thereof.

[0059] One aspect of the present invention provides a stable, ultra-long-acting injectable pharmaceutical composition comprising an insulin analog or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof; collagen; and one or more pharmaceutically acceptable excipients or carriers, wherein the pharmaceutical composition has a pH of 2 to 5; the pharmaceutical composition is a solution upon administration and forms a depot at physiological pH immediately after administration; the pharmaceutical composition exhibits increased bioavailability and a longer duration of action compared to commercially available injectable pharmaceutical compositions containing the "insulin analog or derivative having an isoelectric point of 5 to 8.5" at the same dose; and the "insulin analog or derivative having an isoelectric point of 5 to 8.5" is insulin glargine. The insulin glargine is present in a concentration range of 100 IU to 1,000 IU / ml. In a preferred embodiment of the present invention, the insulin glargine is present in a concentration range of 100 IU to 300 IU / ml.

[0060] One aspect of the present invention provides a stable, ultra-long-acting injectable pharmaceutical composition comprising an insulin analog or derivative having an isoelectric point of 5 to 8.5, or a physiologically acceptable salt thereof; collagen; and one or more pharmaceutically acceptable excipients or carriers, wherein the pharmaceutical composition has a pH of 2 to 5; the pharmaceutical composition is in solution upon administration and forms a depot at physiological pH immediately after administration; the pharmaceutical composition exhibits increased bioavailability and a longer duration of action compared to commercially available injectable pharmaceutical compositions containing the insulin analog or derivative having an isoelectric point of 5 to 8.5 at the same dose; and wherein the collagen present in the composition is human collagen. The human collagen used in the present invention can be obtained by recombinant DNA technology. In a preferred embodiment of the present invention, human collagen type 1 or a rapid human collagen solution prepared by gradually removing salts from an acidic collagen solution using acetic acid and EDTA and then adding acidic water for injection is used. Meanwhile, collagen should be used at a concentration ranging from 3 mg / ml to 10 mg / ml.

[0061] The compositions of the present invention are useful for treating metabolic disorders selected from the group consisting of insulin resistance syndrome, diabetes, hyperlipidemia, fatty liver disease, cachexia, obesity, atherosclerosis, and arteriosclerosis. In a preferred embodiment of the present invention, the compositions of the present invention are useful for treating type 1 and type 2 diabetes.

[0062] The compositions of the present invention exhibit a longer duration of action, i.e., 24 hours or more, when compared to commercially available compositions of the same concentration or dose. The compositions of the present invention may be administered one to three times a week.

[0063] One aspect of the present invention discloses a method for preparing a stable ultra-long acting injectable solution comprising insulin glargine or a physiologically acceptable salt thereof; collagen; and one or more pharmaceutically acceptable excipients or carriers having a pH of 2 to 5, the method comprising the steps of: a) precisely weighing zinc-containing insulin glargine into water for injection and solubilizing it by adding a few μL of 1 M HCl; b) preparing a buffer of pH 2 to 5; c) dissolving an isotonicity agent in a buffer; d) diluting the concentrated insulin glargine of step a) with the buffer of step b); e) adding collagen to the solution of step d) under constant stirring; and f) adjusting the final pH of the solution to between 2 and 5.

[0064] Yet another aspect of the present invention provides a method for modulating the release of an insulin analog, derivative, or metabolite having an isoelectric point of 5.8 to 8.5 from a pharmaceutical composition, comprising formulating insulin glargine with collagen and one or more pharmaceutically acceptable salts at a pH of 2 to 5, and precipitating the composition at physiological pH immediately after administration to create an additional collagen barrier around the insulin. The additional barrier not only controls the release of the insulin glargine precipitate to insulin glargine hexamer and from the hexamer to monomer, but also protects the insulin from protease degradation.

[0065] Each embodiment is further defined in the following examples. The following examples are intended to illustrate the invention, but do not limit the scope of the invention to any formula. [Example]

[0066] Example 1: Preparation of unit composition (100 IU / ml)

[0067] [Table 1]

[0068] Ultra-long acting insulin glargine was produced by first preparing a rapidly polymerizing collagen solution and then using this as an excipient.

[0069] A. Preparation of Rapid Polymerizing Collagen (RPC) Solution A collagen solution was prepared by dissolving 2 g of commercially available recombinant human type 1 collagen in 50 ml of 1 M acetic acid. The collagen solution was dialyzed against a 0.5 M acetic acid + 50 mM EDTA solution and diluted with 0.25 M acetic acid to a collagen concentration of 2 mg / ml. Collagen was precipitated at its isoelectric point (pH 7). The precipitate was washed twice with water for injection. The washed precipitate was suspended in water for injection and dissolved by adding a minimum volume of 1 N HCl. Water for injection was added to dilute the collagen to a concentration of 40 mg / ml. The solution was sterilized and filtered through a 0.2 micron filter. The solution was then placed in a sterilized siliconized glass bottle and stored at 2 to 8°C.

[0070] B. Rapid Polymerization Attribute Evaluation of the Prepared Solution (Polymerization Kinetics of RPC Solution in PBS Environment) 4.5 ml of phosphate buffered saline was dispensed into five glass test tubes, and 0.5 ml of the prepared rapid polymerizing collagen solution was added to each test tube. The polymerization properties were tested and the results were as follows: Time Observation 0 minutes No change 1.0 min Milky white color begins to appear. 5.0 min: Gel with a clear solution interface visible 10 minutes transparent deposit 20 minutes: fibril formation

[0071] The prepared collagen started to form a gel within 1 minute at physiological pH and salt concentration, and formed complete fibrils within 20 minutes in an environment similar to that of subcutaneous tissue.

[0072] C. Manufacture of 5 vials (5,000 IU / 50 ml) of ultra-long-acting insulin glargine (100 IU / ml concentration) 182 mg of insulin glargine (5,000 IU) was suspended in 25 mL of water for injection and dissolved by adding a minimum volume of 1 M HCl. To the insulin glargine solution, 1.5 mg of ZnCl2 (corresponding to Zn), 13.5 mg of m-cresol, 0.1 g of 85% (w / v) glycerol, 0.1 mg of polysorbate 20, and 200 mg of RPC (5.0 mL) were added. A few drops of 1 M HCl were added to the solution to adjust the pH to 4. Water for injection was added to bring the volume to 50 mL. The solution was sterilized and filtered through a sterile 0.2 micron filter. The filtrate was aseptically dispensed into five sterile siliconized glass vials in 10 mL aliquots.

[0073] Example 2: Preparation of a high concentration unit composition (300 IU / ml)

[0074] [Table 2]

[0075] Highly concentrated ultra-long acting insulin glargine could be produced by first preparing a rapidly polymerizing collagen solution and then using this as an excipient.

[0076] A. Preparation of Rapidly Polymerizing Collagen (RPC) Solution: RPC solution was prepared as described in Example 1, Section A.

[0077] B. Preparation of 5 vials (15,000 IU / 50 ml) of ultra-long-acting insulin glargine (300 IU / ml concentration) of Example 2 546 mg of insulin glargine (15,000 IU) was dissolved as described in Example 1B. ZnCl2 (corresponding to 4.5 mg of Zn), 13.5 mg of m-cresol, 0.1 g of 85% glycerol, 0.1 mg of polysorbate 20, and 200 mg of RPC (5.0 ml) were added to the insulin glargine solution. The pH of the solution was adjusted by adding a few drops of 1 M HCl. Water for injection was added to bring the volume to 50 ml. The solution was sterilized and filtered through a sterile 0.2 micron filter. The filtrate was aseptically dispensed into five sterile siliconized glass vials (10 ml each).

[0078] Example 3: Preparation of dilutions for standard curve testing of unit composition (100 IU / ml) and high concentration unit composition (300 IU / ml)

[0079] A. Preparation of Diluted Solutions of Insulin Glargine Composition (100 IU / mL) A diluted solution was prepared by sequentially adding 1.5 mg of ZnCl, 13.5 mg of meta-cresol, 0.1 g of 85% glycerol, 0.1 mg of polysorbate 20, and 200 mg of RPC (5.0 ml) to 25 ml of water for injection. The pH was adjusted to 4 with a minimum of 1 M hydrochloric acid, and then the diluted solution was filled to 50 ml with water for injection. This diluted solution was used as a blank control for insulin glargine injection 100 IU / ml.

[0080] B. Preparation of Diluted Solutions of Insulin Glargine Composition (300 IU / ml) A diluted solution was prepared by sequentially adding 4.5 mg of ZnCl, 13.5 mg of meta-cresol, 0.1 g of 85% glycerol, 0.1 mg of polysorbate 20, and 200 mg of RPC (5 ml) to 25 ml of water for injection. The pH was adjusted to 4 with a minimum of 1 M hydrochloric acid, and then the diluted solution was filled to 50 ml with water for injection. This diluted solution was used as a blank control for insulin glargine injection 300 IU / ml.

[0081] Example 4: Preparation of test solutions and construction of standard curves for HPLC testing The unit composition (insulin glargine solution) of Example 1, which is a super-long-acting insulin glargine solution at 100 IU / ml, was used as a test sample. The unit composition of Example 1 was diluted at different concentrations using the prepared dilution solution. The dilution solution was used as a blank. An aliquot of 2 ml of the diluted sample was placed in a 5 ml 30 Kd cut-off Vivaspin tube. The tube was centrifuged at 5,000 rpm for 30 minutes using a refrigerated centrifuge. The filtrate was used for HPLC analysis as follows.

[0082] <HPLC Test Method> Chromatography system: Agilent HPLC Mode - LC Detector - 214 nm Column - Kromasil C18, 3.0 mm × 250 mm, 4 μ Column temperature - 35 °C Flow rate - 0.6 mL / min Injection volume - 5 μL

[0083]

Table 3

[0084] Analysis - 94.0% to 105.0% on an anhydrous basis.

[0085] <Test Analysis Results>

[0086] The analysis results are as follows, and the standard curve is shown as a graph in Figure 1.

[0087]

Table 4

[0088] Example 5: Development of an in vitro model to simulate the subcutaneous release of insulin glargine Validation of an in vitro test model that mimics the in vivo blood pK profile of insulin glargine requires understanding of the in vivo pK profile of the drug delivered by the subcutaneous ("Sc") route and the in vitro model proposed in the test method.

[0089] Insulin glargine solution is a clear, unbuffered, acidic solution that forms a precipitate after injection into extracellular matrix (ECM) and has an isoelectric point at physiological pH of 7.4. After injection into the body, insulin glargine dissociates into hexamers, which lose Zn and m-cresol upon dilution with surrounding body fluids and are converted into dimers and monomers. The insulin glargine monomers are then metabolized to M1 and M2 in the blood, with M1 being the major metabolite. M1 interacts with insulin receptors in target tissues, exerting its pharmacological effect (hypoglycemic response), and is subsequently degraded by insulin-degrading enzymes into smaller peptides and then amino acids. According to the Tojeo model, 300 IU insulin glargine forms a more concentrated precipitate, resulting in a relatively slower release of the hexamer. This also slows the formation of insulin glargine dimers and monomers. Accordingly, the absorption of the monomer and its conversion to M1 and M2 was observed to be proportionally slower, as shown in Figure 2 below.

[0090] However, no differences in plasma glargine / glargine metabolite pK profiles were observed for identical doses of drug administered by the intravenous route.

[0091] However, all proposed in vitro models must mimic in vivo models, namely, when insulin glargine is delivered via the Sc pathway, it is important to note that the attainment of Cmax during insulin release is proportionally slower with insulin glargine 300 IU / ml than with the 100 IU / ml version at the same dose.

[0092] The inventors conducted a test tube biovalidation of the pK of subcutaneously administered insulin glargine by dividing the test tube into two compartments as shown below. An in vitro test model (see Figure 3) was constructed, and the in vivo reaction was verified by testing the precipitation reaction of insulin glargine in a physiological buffer solution (PBS) dialysis tube representing Sc ECM. The test method was designed so that the insulin glargine precipitate was redissolved in compartment 1 and dissociated into insulin glargine hexamers, dimers, and monomers. In compartment 2, the monomers were absorbed and dispersed through 0.1 μm subcutaneous tissue pores (subcutaneous tissue has capillaries) to form metabolites M1 and M2, which were then dialyzed against the PBS solution in blood through a dialysis membrane. Specifically, compartment 1 corresponds to the redissolution of insulin glargine precipitates in the dialysis tube, and compartment 2 corresponds to the transport / absorption / dispersion of the soluble insulin glargine monomers and the examination of their metabolites. However, if the target tissue is not present or the conditions in compartment 2 in the test tube are not identical to those in the subcutaneous tissue, drug degradation may not occur.

[0093] When the in vitro model of the present invention is treated with glargine at 100 IU / ml (Example 1) or glargine at 300 IU / ml (Example 2), if the observed kinetics are similar to those observed in the in vivo model, it means that the in vitro model of the present invention is suitable for testing the modified release of insulin / insulin glargine dosage forms. Figure 3 shows the concept of the in vitro model proposed in the present invention.

[0094] Example 6: Kinetic study of in vitro protease-induced release of insulin glargine Two-compartment insulin glargine validation devices (in vitro model) were set up as described above (Example 5). Each device contained a 1 L Schott bottle with a wide opening and 100 Kd cutoff dialysis tubing with a 10 mL stopper. Specifically, 5 mL of sterile PBS (pH 7.4) was added to thoroughly pre-cleaned and pre-sterilized dialysis tubing (soaked in 70% aqueous ethanol and equilibrated with PBS). 1 L of sterile PBS was added to a pre-sterilized 1 L bottle, and the dialysis tubing was placed inside. The dialysis tubing was maintained vertically in a floater position. A sterile Teflon-coated 5.08 cm x 0.95 cm magnetic bar was placed in each bottle, and the device was placed on a magnetic stirrer under a sterile hood.

[0095] Set 1 contained 0.6 ml of Lantus solution with an insulin glargine concentration of 100 IU / ml, the same as in Example 1, in PBS solution in dialysis tubing. Set 2 contained 0.2 ml of Tojeo with an insulin glargine concentration of 300 IU / ml, the same as in Example 2.

[0096] To each compartment 1 (dialysis tubing), 10 milliunits of sterilized recombinant rat trypsin and 20 microliters of solution were added. Then, the magnetic stirrer was turned on and the stirring was set to 100 rpm. 0.5 ml samples were collected from each compartment (from the dialysis tubing and bottle) at different intervals. Each sample was analyzed for soluble insulin glargine by HPLC, and the results are shown below.

[0097] [Table 5]

[0098] From the results presented in the table above, it was found that the release of insulin glargine after 30 minutes was similar in both compartments. It was then confirmed that the release of soluble insulin glargine was proportionally delayed in Tojeo (300 IU / ml glargine dosage form) compared to Lantus (100 IU / ml glargine dosage form) due to the formation of more compact insulin glargine precipitate / crystals in compartment 1. This means that our in vitro model experiments confirmed similar observations to in vivo human and animal pK studies.

[0099] Therefore, in the present invention, we decided to use this in vitro method to evaluate the effect of adding rapidly polymerizing collagen (RPC) on the redissolution of glargine from the depot in compartment 1 and the absorption / diffusion of glargine into compartment 2.

[0100] Examples 7 and 8: Examining the effect of different concentrations of collagen on insulin glargine Five 20 ml vials were each filled with 10 ml of insulin glargine at a concentration twice that of 100 IU / ml, and labeled with numbers 1 through 5. Vial 1 was used as a blank, and vials 2 through 5 contained double the amount of rapidly polymerizing collagen (RPC) at 10 mg, 20 mg, 40 mg, and 50 mg (volumes of 0.25 ml, 0.5 ml, 1.0 ml, and 1.25 ml), respectively. Each was then diluted with diluent to a final volume of 20 ml (final insulin glargine concentration of 100 IU / ml).

[0101] Five insulin glargine verification devices (in vitro model) were prepared. 0.5 mL of insulin glargine solution (100 IU / mL) containing 0 mg / mL RPC was placed in compartment 1 (dialysis tubing) as a control solution. Using the same method, 0.5 mL of insulin glargine solution containing 1 mg / mL RPC, 2 mg / mL RPC, 4 mg / mL RPC, and 5 mg / mL RPC was placed in compartment 1 of devices 2 through 5, in the order indicated. 0.5 mL samples were collected from compartment 2 (bottle) of each device at different time intervals, and the samples were analyzed by RP-HPLC to confirm the insulin glargine content.

[0102] [Table 6]

[0103] Meanwhile, 10 ml of insulin glargine was added to each of five 20 ml glass bottles at a concentration twice that of 300 IU / ml, and these bottles were labeled with numbers 1 to 5. Glass bottle 1 was left empty, and glass bottles 2 to 5 were filled with double the amount of rapidly polymerizing collagen (RPC) at 10 mg, 20 mg, 40 mg, and 50 mg (volumes of 0.25 ml, 0.5 ml, 1.0 ml, and 1.25 ml), respectively. Each was then diluted with dilution solution to a final volume of 20 ml (final insulin glargine concentration of 300 IU / ml).

[0104] [Table 7]

[0105] Example 9: In vitro release of the unit composition of Example 1 (100 IU / ml)

[0106] [Table 8]

[0107] Example 10: In vitro release of the unit composition (300 IU / ml) of Example 2

[0108] [Table 9]

Claims

1. A stable, ultra-long-acting injectable pharmaceutical composition comprising insulin glargine having an isoelectric point of 5 to 8.5, a human collagen solution, zinc, cresol, glycerol and polysorbate, The pharmaceutical composition has a pH of 2 to 5; The pharmaceutical composition is in solution upon administration and forms a depot at physiological pH immediately after administration; The pharmaceutical composition is a stable, ultra-long-acting injectable pharmaceutical composition that exhibits increased bioavailability and longer duration of action compared to an injectable pharmaceutical composition containing the "insulin glargine having an isoelectric point of 5 to 8.5" at the same dose.

2. 2. The stable ultra-long acting injectable pharmaceutical composition according to claim 1, wherein the insulin glargine is present in a concentration range of 100 IU to 1,000 IU / ml.

3. The stable, ultra-long-acting injectable pharmaceutical composition according to claim 1, wherein the pharmaceutical composition has a duration of action of 24 hours or more.

4. The stable, ultra-long acting injectable pharmaceutical composition according to claim 1, which is administered two or three times a week.

5. The stable ultra-long-acting injectable pharmaceutical composition according to claim 1, characterized in that the pharmaceutical composition has lower immunogenicity than a commercially available injectable pharmaceutical composition containing the same dose of "insulin glargine having an isoelectric point of 5 to 8.5." 6. A stable, ultra-long acting injectable pharmaceutical composition according to claim 1 for use in treating one or more metabolic disorders.

7. The stable ultra-long acting injectable pharmaceutical composition according to claim 6, characterized in that the metabolic disorder is selected from the group consisting of insulin resistance syndrome, diabetes, hyperlipidemia, fatty liver disease, cachexia, obesity, atherosclerosis and arteriosclerosis.

8. A stable, ultra-long acting injectable pharmaceutical composition as defined in claim 1 for use in a method for treating type 1 or type 2 diabetes in a patient.

9. 1. A method for producing a stable ultra-long acting injectable solution comprising insulin glargine having an isoelectric point of 5 to 8.5, a human collagen solution, zinc, cresol, glycerol and polysorbate, a) preparing an insulin glargine solution by solubilizing insulin glargine with a hydrochloric acid (HCl) solution and water for injection; b) adding the human collagen solution, zinc, cresol, glycerol and polysorbate to the insulin glargine solution of step a) under constant stirring; and c) adjusting the final pH of the solution to 2 to 5.

Images