Compositions and methods of use thereof

Abstract

This disclosure provides a lipid composition and methods of use thereof. This disclosure also provides a lipid composition comprising from about 5-150 mg retatrutide or an equivalent amount of a pharmaceutically acceptable salt thereof, as an active ingredient, one or more pharmaceutically acceptable lipid excipients, and one or more solvents

Description

COMPOSITIONS AND METHODS OF USE THEREOFFIELD

[0001] The present invention relates to lipid compositions comprising retatrutide that can provide an extended duration of action. The present invention also relates to lipid compositions comprising retatrutide, or an equivalent amount of a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable lipid excipients. The present invention also relates to methods and uses of the lipid compositions to treat diabetes, such as type 1 diabetes or type 2 diabetes, obesity, heart failure, obstructive sleep apnea, osteoarthritis, chronic lower back pain, chronic kidney disease, metabolic dysfunction-associated steatotic liver disease (MASLD), the reduction of major adverse cardiac events, and / or the reduction in body weight.BACKGROUND

[0002] Glucose-dependent insulinotropic polypeptide (GIP) receptor and glucagon-like peptide- 1 (GLP-1) receptor active compounds have received much attention and resulted in improved treatments, e.g. in type 2 diabetes and obesity. An example of a GIP / GLP-1 dual agonist is tirzepatide, but many other GIP receptor active compounds are being researched, for example triple agonists of GLP-l / GIP / glucagon receptors are being researched. An example of a triple agonist is retatrutide, developed by Eli Lilly.

[0003] Retatrutide is a single peptide conjugated to a fatty diacid moiety and has agonism toward the GIP, GLP-1, and GCG receptors, herein termed a triple agonist. Retatrutide is less potent at the human GCG and GLP-1 receptors and is more potent at the human GIP receptor, compared to the endogenous receptor ligands. In a phase 2 study (a 48-week, randomized, double-blind, placebo- controlled trial evaluating the efficacy, tolerability, and safety of retatrutide at various doses and dose-escalation regimens in people with obesity, or overweight with weight-related conditions, except type 2 diabetes) retatrutide achieved up to 17.5 % mean weight reduction at 24 weeks in adults with obesity and overweight, as disclosed in a press-release dated June 26, 2023 titled “Lilly's phase 2 retatrutide results published in The New England Journal of Medicine show the investigational molecule achieved up to 17.5% mean weight reduction at 24 weeks in adults with obesity andoverweight”. Although there have been substantial achievements in the treatment of diabetes, obesity etc over the last years more treatments and disease could be targeted with suitable long-acting incretin hormone mimetics.

[0004] Retatrutide also known as LY3437943 has been disclosed e.g. in WO2019 / 125938, which matured in to U.S pat. No.: 11,542,313, both are herein incorporated in their entirety.SUMMARY

[0005] This disclosure provides, in part, a composition comprising a retatrutide, or an equivalent amount of a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable lipid excipients.

[0006] Additionally provided is a composition comprising about 1 wt.% to about 8 wt.% retatrutide or a pharmaceutically acceptable salt thereof, about 30 wt.% to about 55 wt.% diacyl glycerol, about 30 wt.% to about 55 wt.% of phosphatidyl choline, about 5 wt.% to about 40 wt% of a solvent or a solvent mixture.

[0007] Additionally provided is a method of treating a mammalian patient suffering from a disease or condition wherein a first dose of a composition comprising retatrutide or an equivalent amount of a pharmaceutically acceptable salt thereof is administered to the mammalian patient, wherein the composition additionally comprises one or more pharmaceutically acceptable lipid excipients.

[0008] Additionally provided is a method of treating a mammalian patient suffering from a disease or condition wherein composition comprising about 1 wt.% to about 8 wt.% retatrutide or a pharmaceutically acceptable salt thereof, about 30 wt.% to about 55 wt.% diacyl glycerol, about 30 wt.% to about 55 wt.% of phosphatidyl choline, about 5 wt.% to about 40 wt% of a solvent or a solvent mixture is administered to the mammalian patient.DETAILED DESCRIPTION

[0009] The features and other details of the disclosure will now be more particularly described. Before further description of the present disclosure, certain terms employed in the specification, examples and appended claims are collected here. These definitions should be read in light of the remainder of the disclosure and understood as by a person of ordinary skill in the art. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art.BRIEF DESCRIPTION OF THE DRAWINGS

[0010] FIG. 1 illustrates the pharmacokinetic profile (0-672 h), of retatrutide in a composition of the present disclosure.DEFINITIONS

[0011] “Treating” includes any effect, e.g., lessening, reducing, modulating, or eliminating, that results in the improvement of the condition, disease, disorder, and the like.

[0012] ‘ ‘Individual,” “patient,” or “subject” are used interchangeably and include any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, swine, cattle, sheep, horses, or primates, and most preferably humans.

[0013] The term “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” as used herein refers to any and all solvents, dispersion media, coatings, isotonic and absorption delaying agents, and the like, that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art.

[0014] The term “pharmaceutical composition” as used herein refers to a composition comprising at least one compound as disclosed herein formulated together with one or more pharmaceutically acceptable carriers.

[0015] In the present specification, the term “therapeutically effective amount” means the amount of the subject compound that will elicit the biological or medical response of a tissue, system, animal or human that is being sought by the researcher, veterinarian, medical doctor or other clinician. The compounds of the disclosure are administered in therapeutically effective amounts to treat a disease. Alternatively, a therapeutically effective amount of a compound is the quantity required to achieve a desired therapeutic and / or prophylactic effect, such as an amount which results in the treatment of one or more of the diseases listed herein.

[0016] Steady state is the time during which the concentration remains stable or consistent when the drug is given repeatedly. The steady state for the administration of the GIP receptor active compounds is estimated, e.g. in healthy volunteers to be in the range of 2-3 months, such as about 2 months.

[0017] All % are specified by weight herein throughout, unless otherwise indicated. Percent (%) by weight may be abbreviated e.g., as wt.%. Furthermore, the % by weight indicated is the % of the total lipid composition including all the components indicated herein, unless otherwise indicated.

[0018] Whenever a range is used, for example in the list of embodiments, it is to be understood that in the event a range of for example 8- 10_3 is disclosed it is intended to be understood as each individual listing are explicitly included. In the example of 8- 10_3 it would mean 8, 9, 10, 10_l, 10_2 and 10_3 are considered explicitly included in the range.Methods

[0019] The disclosure provides, in part, method of treating a disease or condition is selected from the group consisting of Alzheimer's disease, hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive disorders, heart failure, obstructive sleep apnea, osteoarthritis, chronic lower back pain, chronic kidney disease, metabolic dysfunction- associated steatotic liver disease (MASLD), the reduction of major adverse cardiac events, the reduction in body weight, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia, Parkinson's disease, polycystic ovary syndrome, MASH (metabolic dysfunction-associated steatohepatitis) and gastric ulcers lipid compositions comprising administering retatrutide, or an equivalent amount of a pharmaceutically acceptable salt thereof to a patient. In some embodiments the administration, e.g. the maintenance administration, of retatrutide, or pharmaceutically acceptable salt thereof is once every two to five weeks, such as once every 14 days ± 4 days, such as once every 28 days ± 7 days, such as once every 28 days ± 6 days, such as once every 28 days ± 5 days, such as once every 28 days ± 4 days, such as once every 28 days ± 3 days, such as once every 28 days ± 2 days, such as once every 28 days ± 1 days.In some embodiments the administration is once every four weeks, such as once every 28 days ± 3 days, such as once every 28 days ± 2 days, such as once every 28 days ± 1 days, such as once every 28 days. In an embodiment the method is for treating diabetes, obesity, heart failure, obstructive sleep apnea, osteoarthritis, chronic lower back pain, chronic kidney disease, metabolic dysfunction-associated steatotic liver disease (MASLD), the reduction of major adverse cardiac events, and / or the reduction in body weight.Compounds

[0020] The active agent in the present disclosure is retatrutide, a GIP receptor, a GLP-1 receptor and a glucagon receptor agonist (triple agonist). Retatrutide, also known as LY3437943, has been disclosed e.g. in WO2019 / 125938, which matured into U.S pat. No.: 11,542,313, and the CAS number for retatrutide is 2381089-83-2. Retatrutide is an experimental drug developed by Eli Lilly for treating obesity, type 2 diabetes, and non-alcoholic fatty liver disease (NAFLD). It works by targeting three hormone receptors: GLP-1, GIP, and glucagon.

[0021] The compositions disclosed herein comprise from about 2 mg to about 80 mg of retatrutide, or an equivalent amount of a pharmaceutically acceptable salt thereof, and is generally made up from about 10 to about 99 wt% lipids, e.g. about 15 to about 99 wt% lipids, e.g. about 30 to about 90 wt% lipids, e.g. about 35 to about 85 wt% lipids, about 40 to about 85 wt% lipids, about 50 to about 85 wt% lipids, about 60 to about 85 wt% lipids, e.g. about 70 to about 85 wt% lipids. Ratios of lipids, may in the case of GDO : PC be 30:70 to 60:40 or 40:60 to 70:30 (e.g., 45:55 to 55:45, 40:60 to 54:46, 45:55 to 54:46, 47:53 to 53:47), or ratios of around 50:50 (e.g., 49:51 to 51:49).

[0022] Example of lipids making up the lipid compositions disclosed herein are diacyl glycerol (e.g. glycerol dioleate) and phosphatidylcholine (e.g. soy phosphatidylcholine). The composition comprises a solvent or a mixture of solvents, examples are ethanol, propylene glycol (PG), dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), and glycerol. The solvent(s) may be present in a total amount of about 0.5 - about 40 wt%, e.g. about 1 - about 30 wt%, e.g. about 2 - about 25 wt%, e.g. about 8 - about 25 wt%, e.g. about 1 0 - about 25 wt%, e.g. about 12 - about 22 wt%, e.g. about 3 - about 20 wt%, e.g. about 4 - about 19 wt%, e.g. about 4 - about 18 wt%. e.g. about 4 - about 17 wt%, e.g. about 4 - about 16 wt%, e.g. about 4 - about 15 wt%, e.g. about 4 - about 14 wt%, e.g. about 4 - about 13 wt%, e.g. about 4 - about 12 wt%, e.g. about 4 - about 11 wt%, e.g. about 4 - about 10 wt% of the total composition. The lipid composition optionally includes additional excipients such as stabilizing agents, antioxidants etc. The composition may contain water, although in some aspect of the present disclosure the water content of the lipid composition at release of the product is not more than (NMT) 2%, such as less than 1.5 wt.% determined by USP<921>. The inactive excipients, i.e., the lipids, such as diacyl glycerol, e.g. glycerol dioleate and phosphatidyl choline, the solvent and / or solvent mixture and any optional additional excipients adds up to at least 70 wt.%, such as 75 wt.%, such as at least 80 wt.%, such as at least 85 wt.% such as at least 90 wt.% of the total drug product composition. The rest is the retatrutideand by-products, for example, by-products present in the lipids. Glycerol dioleate (GDO) may also contain monoglycerides (NMT 2%) and triglycerides (NMT 5%). Generally, the GDO used in the lipid composition should contain at least about 93% glycerol dioleate. Phosphatidyl choline may also contain lysophosphatidylcholine (NMT 3%) and triglycerides (NMT 2%). Generally, the phosphatidyl choline used in the lipid composition should contain at least about 94% phosphatidyl choline.

[0023] The lipid composition is in some embodiments provided in a pre-filled syringe, e.g., a glass syringe, equipped with a needle and the drug product is not necessary to be reconstituted and considered ready-to-use. Suitable syringes may be those having a volume of about 0.5, about 0.75, about 1.0, about 1.25, and about 1.5 mL. One example is a 1 mL glass syringe, optionally equipped with a needle, e.g. 23G, 25G, 27G or 29G needle. An additional example is a suitable auto-injector, e.g. an auto-injector suitable for a glass syringe or glass cartridge. An additional example is a pen injector.

[0024] In many aspects of the present disclosure the lipid composition is substantially non-aqueous, e.g. it contains less than 3 wt% water, such as less than 2 wt% water, such as less than 1 wt% water.

[0025] The lipid compositions may form a depot composition upon contact with an aqueous fluid. The term “depot” relates to the composition which is formed upon exposure of the lipid composition to excess aqueous fluid, e.g., as occurs during numerous parenteral administration routes, such as administration in the subcutaneous tissue of a human patient. The depot typically has a much higher viscosity than the corresponding lipid composition and provides for the gradual release of the retatrutide.Lipid Composition

[0026] This disclosure provides, in part, a lipid composition comprising from about 2 mg to about 80 mg of retatrutide, or an equivalent amount of a pharmaceutically acceptable salt thereof, as an active ingredient, one or more pharmaceutically acceptable lipid excipients, and one or more solvents.

[0027] The lipid composition may form a depot composition upon contact with an aqueous fluid. The term “depot” relates to the composition which is formed upon exposure of the lipid composition to excess aqueous fluid, e.g., as occurs during numerous parenteral administration routes, such as administration in the subcutaneous tissue of a human patient. The depot typically has a higher viscosity than the corresponding lipid composition and provides for the gradual release of retatrutide or a pharmaceutically acceptable salt thereof contained within the depot. The lipid composition is intended for pharmaceutical use.Pharmaceutically Acceptable Lipid Excipients

[0028] The lipid composition is made up from about 10 to about 99 wt.% (e.g., about 15 to about 99 wt.%, about 30 to about 90 wt.%, about 40 to about 90 wt.%, about 40 to about 85 wt.%, about 50 to about 85 wt.%, about 60 to about 85 wt.%, about 70 to about 85 wt.%) pharmaceutically acceptable lipid excipients.

[0029] Example of pharmaceutically acceptable lipid excipients making up the lipid compositions disclosed herein are diacyl glycerols (e.g., glycerol dioleate (GDO)) and phosphatidylcholine (PC) (e.g., soy phosphatidylcholine).

[0030] Ratios of lipids may in the case of GDO: PC be 30:70 to 60:40 or 40:60 to 70:30 (e.g., 45:55 to 55:45, 40:60 to 54:46, 45:55 to 54:46, 47:53 to 53:47), or ratios of around 50:50 (e.g., 49:51 to 51:49). Diacyl glycerol

[0031] In one embodiment, the lipid composition contains diacyl glycerol and a suitable example thereof is glycerol dioleate (GDO). The GDO may be present in the lipid composition in an amount ranging from 20 to 60 wt.% of the lipid composition (e.g., about 30 to about 50 wt.%, about 32 to about 48 wt.%, about 33 to about 45 wt.%, about 34 to about 42 wt.%). Additional examples of GDO in the compositions disclosed herein are about 30 to about 48 wt.%, e.g. about 30 to about 43 wt.%, e.g. about 33 to about 42 wt.%.

[0032] Suitable diacyl glycerols, such as GDO, may be derived from natural sources, there is generally a certain proportion of “contaminant” lipid having other chain lengths, etc. In this context, “pure” GDO is a di-ester of glycerol and two Cl 8:1 fatty acids. Any other diacyl glycerol is considered to be an impurity. In one aspect, GDO, as used herein, indicates any commercial grade of GDO with concomitant impurities (i.e., GDO of commercial purity). These impurities may be separated and removed by purification but, providing the grade is consistent, this is rarely necessary. If necessary, however, “GDO"” may be essentially chemically pure GDO, such as at least 70% pure GDO (e.g., at least 75% pure, at least 80% pure, at least 85% pure, at least 90% pure, at least 93%, at least 95% pure, at least 98% pure, at least 99% pure (area%)). In some embodiments, the diglycerides content of the glycerol dioleate is about 92% - 100%, such as about 93% to about 99%. The GDO used herein should have an oleic acid content (C18:l) of at least 80% (e.g., at least 85%, at least 90%, at least 95%, at least 98%, at least 99%). In some embodiments, the diglyceride content is about 93% to about 99%, and the oleic acid content is about 88% to about 99% (area%). Diacyl glycerol other than GDO, or mixtures of GDO and other diacyl glycerols are also useful.

[0033] Any material used in the lipid composition, including GDO, may potentially include unavoidable trace impurities of metals, optionally including heavy metals. A typical maximum concentration of heavy metals (or elemental impurities) in diacyl glycerol, e.g., GDO, is 5 ppm.

[0034] In alternative embodiments, GDO may be replaced by, or combined with, at least one other lipid, e.g., at least one fatty acid and / or fatty acid ester (lipid). Fatty acids / lipids contain a polar carboxylic acid or ester “head group” with the hydrocarbon chain forming a non-polar “tail” group. Fatty acid esters are esterified fatty acids. Fatty acids or esters used in the lipid composition may be solid or liquid at room temperature and pressure. Examples of non-polar “tail” groups include C6-C32 alkyl and alkenyl groups, which are typically present as long chain carboxylic acids or the esters thereof. These are often described by reference to the number of carbon atoms and the number of unsaturations in the carbon chain. Thus, CX:Z indicates a hydrocarbon chain having X carbon atoms and Z unsaturations. Examples particularly include caproyl (C6:0), capryloyl (C8:0), capryloyl (C10:0), lauroyl (C12:0), myristoyl (C14:0), palmitoyl (C16:0), phytanoyl (C16:0), palmitoleoyl (C16:l), stearoyl (C18:0), oleoyl (C18:l), elaidoyl (C18:l), linoleoyl (C18:2), linolenoyl (C18:3), arachidonoyl (C20:4), behenoyl (C22:0), and lignoceroyl (C24:9) groups. When reference is made herein to the number of carbon atoms in the “chain” or “tail” this number includes the carbon atom of the -C(O)O-moiety, as is conventional in the art.

[0035] Thus, typical non-polar chains are based on the fatty acids of natural ester lipids, including caproic, caprylic, capric, lauric, myristic, palmitic, phytanic, palmitolic, stearic, oleic, elaidic, linoleic, linolenic, arachidonic, behenic, or lignoceric acids, or the corresponding alcohols.

[0036] The lipid(s) may be saturated or unsaturated. For example, the lipid(s) may comprise at least 1 wt.% unsaturated lipid (based on the total lipid content) (e.g., at least 5 wt.% (5-100%), at least 15 wt.% (15-100%), at least 30 wt.% (30-100%), at least 50 wt.% (50-100%), at least 80 wt.% (80-100%)).

[0037] In other alternative embodiments, GDO may be replaced or combined with, for example, an edible lipid such as almond oil, avocado oil, butter, canola oil, castor oil, coconut oil, corn oil, cottonseed oil, flaxseed oil, ghee, lard, linseed oil, macadamia oil, margarine, mustard oil, olive oil, palm oil, peanut oil, pumpkin seed oil, rice bran oil, safflower oil, sesame oil, soybean oil, sunflower oil, tea seed oil, vegetable oil, or walnut oil.

[0038] One particular example is tocopherol, which may be used to replace GDO, or tocopherol may be used in combination with GDO, e.g., in a ratio of 30:70; 40:60 or 50:50.

[0039] When GDO is replaced by at least one other lipid, the at least one lipid may be present in the lipid composition in an amount ranging from 20 to 90 wt.% of the lipid composition (e.g., 30 to 70 wt.%,33 to 60 wt.%, 43 to 60 wt.%, 38 to 43 wt.%). If the lipid composition contains both GDO and at least one additional lipid, the combination may also be present in the lipid composition in an amount ranging from 20 to 90 wt.% of the lipid composition (e.g., 30 to 70 wt.%, 33 to 60 wt.%, 43 to 60 wt.%, 38 to 43 wt.%). Phosphatidyl Choline (PC)

[0040] The lipid composition typically also contains phosphatidyl choline (PC). When present, PC is present in an amount ranging from 20 to 60 wt.% of the lipid composition (e.g., about 30 to about 50 wt.%, about 32 to about 48 wt.%, about 33 to about 45 wt.%, about 34 to about 42 wt.%). Additional examples of PC in the compositions disclosed herein are about 30 to about 48 wt.%, e.g. about 30 to about 43 wt.%, e.g. about 33 to about 42 wt.%. Ratios of GDO : PC may be 30:70 to 40:60 be 30:70 to 60:40 or 40:60 to 70:30 (e.g., 55:45 to 45:55, 45:55 to 55:45, 40:60 to 54:46, 45:55 to 54:46, 47:53 to 53:47). Ratios of around 50:50 (e.g., 49:51 to 51:49 or 50:50 to 48:52) may be highly effective.

[0041] In alternative embodiments, PC may be replaced by, or combined with, at least one other phospholipid. The phospholipids may be derived from a natural source. In the case of PC, suitable sources of phospholipids include egg, heart (e.g., bovine), brain, liver (e.g., bovine), and plant sources, including vegetable PC, such as soybean. Such sources may provide one or more constituents of the PC and / or at least one other phospholipid. Any single PC or mixture thereof may be used according to the present disclosure, although egg, heart (e.g., bovine), brain, liver (e.g., bovine), and plant sources, including vegetable PC, such as soybean or any mixture thereof are used in most embodiments disclosed herein.

[0042] The PC may be derived from soy. The PC may comprise 18:2 fatty acids as the primary fatty acid component with 16:0 and / or 18:1 as the secondary fatty acid components. The fatty acid components may be present in the PC at a ratio of between 1.5: 1 and 6:1. PC having approximately 60-65% 18:2, 10 to 20% 16:0, 5-15% 18:1, with the balance predominantly other 16 carbon and 18 carbon fatty acids may be used, e.g., soy PC. In some embodiments, the PC has a purity of not less than 90%, not less than 92%, not less than 94%, such as about 94% - 100% (based on dry weight). The maximum lysophosphatidylcholine content is about 3%, e.g., not more than 3%. The maximum triglycerides content is about 2%, such as not more than 2%. In some embodiments, the PC used in the lipid composition has a purity of about 94% - 100% (based on dry weight), and contains not more than 3% lysophosphatidylcholine and optionally not more than 2% triglycerides.

[0043] Alternatively, the PC component may comprise synthetic dioleoyl PC (DOPC). The use of DOPC may provide increased stability and so may be used for compositions needing to be stable to long term storage, and / or having a long release period in vivo. Here, the PC component may contain at least50% synthetic dioleoyl PC (e.g., at least 75% synthetic dioleoyl PC) and even essentially pure synthetic dioleoyl PC. Any remaining PC can be of any sort. DOPC may be more expensive.

[0044] Alternative types of phospholipids are synthetic or highly purified PCs, such as dioleoyl phosphatidyl choline (DOPC), and may, in alternative embodiments, be used as all or part of the phospholipid component. The synthetic dioleoyl PC may be l,2-dioleoyl-sn-glycero-3-phosphocholine, and other synthetic PC components include DDPC (l,2-Didecanoyl-sn-glycero-3-phosphocholine); DEPC (l,2-Dierucoyl-sn-glycero-3-phosphocholine); DLOPC (l,2-Dilinoleoyl-sn-glycero-3- phosphocholine); DLPC (l,2-Dilauroyl-sn-glycero-3-phosphocholine); DMPC (1,2-Dimyristoyl-sn- glycero-3-phosphocholine); DOPC (l,2-Dioleoyl-sn-glycero-3-phosphocholine); DPPC (1,2- Dipalmitoyl-sn-glycero-3-phosphocholine); DSPC (l,2-Distearoyl-sn-glycero-3-phosphocholine); MPPC (l-Myristoyl-2-palmitoyl-sn-glycero 3-phosphocholine); MSPC (l-Myristoyl-2-stearoyl-sn-glycero-3- phosphocholine); PMPC (l-Palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine); POPC (l-Palmitoyl-2- oleoyl-sn-glycero-3-phosphocholine); PSPC (l-Palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine); SMPC (l-Stearoyl-2-myristoyl-sn-glycero-3-phosphocholine); SOPC (l-Stearoyl-2-oleoyl-sn-glycero-3- phosphocholine); and SPPC (l-Stearoyl-2-palmitoyl-sn-glycero-3-phosphocholine), or any combination thereof.

[0045] Alternative lipid compositions that may be used in the present disclosure are disclosed in WO 2016 / 066655, which is incorporated herein by reference, where lipid slow-release matrices based on triacyl lipids that can form depot compositions on exposure to aqueous fluids without the need for a phospholipid component to be present, though in certain embodiments a phospholipid may also be present.

[0046] Alternative embodiments include phosphatidylethanolamine (PE), phosphatidylserine, and phosphatidylinositol, instead of phosphatidyl choline.

[0047] Since the lipid compositions are to be administered to a subject, such as a patient, including retatrutide, the components should be biocompatible. In this regard, PC (such as soy PC and / or DOPC) and GDO are useful, since they are well tolerated and are broken down in vivo into components that are naturally present in the mammalian body. Appropriate amounts of each component suitable for the combination are those amounts indicated herein for the individual components in any combination. This applies also to any combinations of components indicated herein, where context allows.

[0048] When PC is replaced by at least one other phospholipid, the at least one phospholipid may be present in the lipid composition in an amount ranging from 20 to 80 wt.% of the lipid composition (e.g., 30 to 70 wt.%, 33 to 55 wt.%, 35 to 55 wt.%, 38 to 43 wt.%). If the lipid composition contains both PCand at least one additional phospholipid, the combination may also be present in the lipid composition in an amount ranging from 20 to 80 wt.% of the lipid composition e.g., 30 to 50 wt.%, 33 to 45 wt.%, 34 to 44 wt.%, 35 to 43 wt.%). Ratios of GDO and / or the at least one lipid other than GDO : PC and / or the at least one phospholipid other than PC may be 30:70 to 60:40 or 40:60 to 70:30 (e.g., 55:45 to 45:55, 45:55 to 55:45, 40:60 to 54:46, 45:55 to 54:46, 47:53 to 53:47). Ratios of around 50:50 (e.g., 49:51 to 51:49) may be highly effective.

[0049] The inactive excipients, i.e., the lipids, such as diacyl glycerol, e.g., glycerol dioleate and phosphatidyl choline, the solvent and / or solvent mixture, and any optional additional excipients add up to at least 70 wt.%, such as 75 wt.%, such as at least 80 wt.%, such as at least 85 wt.%, such as at least 90 wt.% of the total drug product lipid composition. The rest is Retatrutide and by-products, for example, originating from the lipids. Glycerol dioleate (GDO) may also contain monoglycerides (NMT 2%) and triglycerides (NMT 5%). Generally, the GDO used in the lipid composition should contain at least about 93% glycerol dioleate. Phosphatidyl choline may also contain lysophosphatidylcholine (NMT 3%) and triglycerides (NMT 2%). Generally, the phosphatidyl choline used in the lipid composition should contain at least about 94% phosphatidyl choline.Solvent

[0050] The lipid composition also contains one or more solvents, e.g., ethanol, propylene glycol, dimethyl sulfoxide, N-methyl-2-pyrrolidone, and glycerol or a mixture thereof. In many embodiments, ethanol is a suitable solvent. Ethanol may be replaced by, or combined with, at least one other biocompatible organic solvent. Since the lipid composition may generate a depot composition following administration (e.g., in vivo), typically upon contact with excess aqueous fluid, it is desirable that this solvent be tolerable to the subject and be capable of mixing with the aqueous fluid, e.g., body fluids, and / or diffusing or dissolving out of the lipid composition into the aqueous fluid. Therefore, solvents having at least moderate water solubility may be used. In some embodiments the lipid compositions include a polar solvent. Typically, a level of 1 to 30 wt.% (e.g., 2 to 25 wt.%, 2 to 20 wt.%, 2 to 18 wt.%, 2 to 16 wt.%, 2 to 15 wt.%, 4 to 15 wt.%, 5 to 15 wt.%,) solvent will provide suitable release and viscosity properties.

[0051] The solvent(s) may be present in a total amount of about 0.5 - about 40 wt.% (e.g., about 1 - about 30 wt.%, about 2 - about 25 wt.%, about 8 - about 25 wt.%, about 10 - about 25 wt.%, about 12 - about 22 wt.%, about 3 - about 20 wt.%, about 4 - about 19 wt.%, about 4 - about 18 wt.%. about 4 - about 17 wt.%, about 4 - about 16 wt.%, about 4 - about 15 wt.%, about 4 - about 14 wt.%, about 4 - about 13 wt.%, about 4 - about 12 wt.%, about 4 - about 11 wt.%, about 4 - about 10 wt.%) of the totallipid composition. In some embodiments, the total amount of solvent is at a level of 5 - 40 wt.%, e.g., 10- 30 wt.%, 9-25 wt.%, such as 10- 25 wt.%, e.g. 10-20 wt.%. or 8-18 wt.%.

[0052] As indicated above, the amount of solvent in the lipid compositions may be at least sufficient to provide a low viscosity mixture (e.g., a molecular solution) of the components of the lipid composition and can be determined for any particular combination of components by standard methods.

[0053] The solvent may be a single solvent (e.g., ethanol, DMSO, and NMP) or a mixture of suitable solvents (e.g., ethanol and PG) but will generally be of low viscosity. In some embodiments the solvent is a mixture of ethanol and propylene glycol. In some embodiments the solvent is a mixture of ethanol and DMSO. In some embodiments the solvent is a mixture of ethanol and propylene glycol and glycerol. In some embodiments the solvent is a mixture of DMSO and ethanol and / or glycerol. The viscosity of the “low viscosity” solvent (single solvent or mixture) may be no more than 18 mPas at 20°C (e.g., no more than 15 mPas at 20°C, no more than 10 mPas at 20°C, no more than 7 mPas at 20°C).

[0054] WO 2012 / 160213, herein incorporated by reference, describes the addition of a polar solvent in addition to a mono-alcoholic solvent results in numerous advantages including reduced viscosity and reduced active agent burst profile. In addition to the aspects described previously for the solvent component, in some embodiments, the solvent comprises a mono-alcoholic solvent (e.g., ethanol) and a polar co-solvent. The term “polar co-solvent” or “polar solvent” as used herein defines a solvent having a dielectric constant (diel) of at least 28 at 25°C (e.g., at least 30 at 25°C) but is not water or any aqueous fluid. Examples include dimethyl sulfoxide (diel about 47), propylene glycol (diel about 32), and N- methyl-2-pyrrolidone (NMP, diel about 32). The levels of solvent recited herein may apply equally to mixtures of mono-alcoholic solvent and a polar co-solvent unless context permits otherwise.

[0055] In another embodiment, the solvent comprises, consists essentially of, or consists of a mixture of a mono-alcoholic solvent and a co-solvent. The co-solvent may be a di-alcoholic C3-C6 organic solvent, i.e., a C3-C6 organic solvent comprising two hydroxy groups, or another solvent. The di-alcoholic solvent may be propylene glycol. Another co-solvent may be DMSO. When present, a co-solvent may be included at a level of 1 to 12 wt.% of the lipid composition (e.g., 3 to 10 wt.%, 4 to 9 wt.%). This level is counted as part of the ranges recited above for the solvent. In one embodiment, the solvent comprises, consists essentially of, or consists of a mixture of ethanol and propylene glycol (PG) and / or DMSO.

[0056] Where both an organic mono-alcoholic solvent and a polar co-solvent are present, e.g., ethanol and PG and / or DMSO, the amounts and ratio of ethanol and PG and / or DMSO may have an effect on properties such as release of an active agent, viscosity of the lipid composition, etc., features which are allimportant characteristics of suitable lipid compositions for subcutaneous injection of an active agent to a patient in need thereof. Such ratio and amounts are contemplated and disclosed herein.

[0057] In one embodiment, the total amount of solvent is at a level of 5 to 35 wt.% (e.g., 7 to 25 wt.%, 10 to 23 wt.%, 10 to 20 wt.%) and comprises, consists essentially of, or consists of a mixture of ethanol and DMSO, wherein the ratio of ethanol to DMSO (w / w) is in the range of 40:60 to 95:5 (w / w)). Additional examples are about 6-15 wt.% ethanol and about 1-10 wt.% DMSO, e.g., about 8-12 wt.% ethanol and 2-8 wt.% of DMSO.

[0058] In one embodiment, the solvent is dimethyl sulfoxide (DSMO), which may be present at a level of 1 to 30 wt.% (e.g., 2 to 15 wt.%, 3 to 12 wt.%, 2 to 10 wt.%). Additional examples are about 2-15 wt.% DMSO and 2-8 wt.% DMSO.

[0059] In an embodiment the solvent is selected from the group consisting of ethanol, propylene glycol, dimethyl sulfoxide, N-methyl-2-pyrrolidone, and glycerol or a mixture thereof.

[0060] In an embodiment the solvent is selected from the group consisting of ethanol, and / or propylene glycol, and / or dimethyl sulfoxide.

[0061] In an embodiment the solvent is selected from the group consisting of ethanol, ethanol / PG, ethanol / glycerol, ethanol / DMSO, ethanol / DMSO / glycerol, and dimethyl sulfoxide.

[0062] Even where a polar solvent is present in the lipid composition, the total water level may remain at release of the lipid composition for sale at the levels described herein (e.g., 1 wt.% or less, 0.1 to 1.0 wt.%).

[0063] The term “release” in this context means released for administration to a patient, i.e., a pharmaceutical product approved by a regulatory agency, e.g., the FDA. The term release specification means the tests and limits against which raw material, intermediate, and final product are measured prior to use and / or release.

[0064] In some embodiments, the lipid composition contains water, e.g., as a solvent. In other embodiments, the lipid composition is substantially non-aqueous, e.g., it contains less than 3 wt.% water, such as less than 2 wt.% water, such as less than 1 wt.% water. In other embodiments, the water content of the lipid composition at release of the product is not more than (NMT) 1 wt.%, such as 0.5 wt.%, such as 0.1 wt.%, determined by USP<921>.Water content

[0065] It is difficult to eliminate all traces of water (especially from the raw materials). Even if essentially water-free formulations could be achieved, lipid compositions described in this disclosure willtypically be stored in ready-to-use form, e.g., in syringes and possibly under refrigerated conditions, or alternatively frozen conditions. Syringes are often not completely air-tight meaning that the level of water in the lipid composition may increase to an appreciable level over time, e.g., over months, even if the initial level of water is insignificant.

[0066] The initial absolute level of water in the lipid composition disclosed herein may be between 0 to 3 wt.% (e.g., less than 2 wt.%, less than 1.5 wt.%, less than 1 wt.%, less than 0.5 wt.%, less than 0.1 wt.%). For example, the level of water may be in the range of 0.1 to 0.9 wt.%, such as 0.2 to 0.8 wt.%, such as 0.3 to 0.7 wt.%, such as 0.4 to 0.6 wt.%. These levels refer to the absolute level of water and not added levels of water. Any unavoidable trace of water present within the components of the lipid composition is included in this stated level of water. After 3 months of storage, the absolute water level may be no more than 1.5 wt.%. Absolute levels of water can be measured by methods well known in the art, such as Karl Fischer titration. For example, the water content may be measured according to the procedure in United States Pharmacopoeia (USP 40 - NF 35, USP <921> Water determination, Method la).

[0067] The lipid composition disclosed herein has a water content of between 0 to 2 wt.% (e.g., less than 1.5 wt.%, less than 1 wt.%, less than 0.5 wt.%) when the lipid composition product is released for sale (e.g., released for administration to a patient).

[0068] In some embodiments the lipid composition disclosed herein is intended for refrigerated storage, e.g., in order to achieve suitable stability the lipid composition comprising retatrutide or an equivalent amount of a pharmaceutically acceptable salt thereof, e.g., a lipid composition as disclosed herein provided in a syringe, such as a glass syringe, or a glass container for an injection device, such as an auto-injector is stored at refrigerated conditions, or alternatively frozen conditions. In some aspect the stability of the lipid composition comprising retatrutide or an equivalent amount of a pharmaceutically acceptable salt thereof fulfilling regulatory stability requirements for at least six months, such as at least 12 months, such as at least 18 months upon storage at 2-8°C (refrigerated conditions).Optional excipients

[0069] Besides retatrutide or a pharmaceutically acceptable salt thereof, one or more lipids, and solvent(s), the lipid composition may contain additional excipient(s), such as stabilizing agents, antioxidants, etc.

[0070] Retatrutide, or pharmaceutically acceptable salt thereof, in the lipid compositions disclosed herein, may gain stability (both storage and in vivo stability) by certain stabilizing additives. Suchadditives include, but are not limited to, sugars (e.g., sucrose, trehalose, lactose, etc.), polymers (e.g., polyols such as carboxy methyl cellulose), amino acids (such as methionine, glutamate, lysine, etc.), lipid- soluble acid components such as HC1, anionic lipids, and / or surface active agents (such as dioleoyl phosphatidyl glycerol (DOPG), palmitoyloleoyl phosphatidylglycerol (POPG), and oleic acid (OA)). Another example is EDTA (“ethylenediamine tetraacetic acid” or “edetic acid”). As used herein, the term “EDTA” may represent ethylenediammetetraacetic acid as such. Alternatively, EDTA as indicated herein may include ethylenediammetetraacetic acid itself, EDTA analogues, and alkylammonium EDTA salts. “EDTA” herein thus includes “EDTA, analogues thereof, and alkylammonium EDTA salts,” whenever context allows.

[0071] Examples of EDTA analogues include:Iminodiacetic acid (IDA) - (NH(CH2CO2H)2;Nitrilotriacetic acid (NTA) - N(CH2CO2H)3;Pentetic acid* - N(CH2CO2H)2CH2CH2N(CH2CO2H)CH2CH2N(CH2CO2H)2;Egtazic acid - N(CH2CO2H)2CH2CH2OCH2CH2OCH2CH2N(CH2CO2H)2;NOTA - [N(CH2CO2H)CH2CH2]3; andDOTA - [N(CH2CO2H)CH2CH2]4.* Also known as “DTPA”

[0072] EDTA analogues and alkylammonium salts thereof are further disclosed and described in WO 2018 / 060212, which is hereby incorporated herein by reference.

[0073] The alkylammonium salt(s) of EDTA is provided by contacting the EDTA or analogue thereof with a suitable alkylamine, examples are:Ethanolamine “ETA” (NH2(CH2CH2OH));Diethanolamine “DiETA” (NH(CH2CH2OH)2); meglumine (NH(CH3)CH2(CHOH)4CH2OH)) ; tris-hydroxymethylamine “TRIS” (N(CH2OH)3); ethylenediamine (NH2CH2CH2NH2); or serinol (NH2CH(CH2OH)2).

[0074] An example of alkylammonium EDTA salt is EDTA with ethanolamine (ETA) (e.g., EDTA with ETA only).

[0075] The lipid composition may also contain EDTA salts comprising an anion of EDTA and at least one alkylammonium cation of the suitable alkylamine as previously described. Further examples and details are available in WO 2018 / 060212, herein incorporated by reference.

[0076] Single-dose formats, such as pre-filled syringes comprising lipid compositions disclosed herein, must remain stable and potent in storage prior to use but are disposable after the single use. The substantially non-aqueous lipid compositions have sufficient storage stability at elevated temperatures, such as at 25 °C or even 40°C.

[0077] A single dose format of the lipid composition may have a stability such that after storage for 2 months at 25°C , the assayed concentration is at least 85% that of the initial assayed Retatrutide concentration, and after 3 months, the assayed concentration is at least 80% that of the initial assayed concentration.

[0078] The lipid compositions disclosed herein can optionally contain an antimicrobial or microbial- static agent, which includes bacteriostatic agents and preservatives. Such agents include benzalkonium chloride, m-cresol, benzyl alcohol, or other phenolic preservatives. Typical concentrations as known in the art can be used. In most aspects and embodiments, there are no antimicrobial or microbial-static agents added.

[0079] These additional components, if present, may be present in an amount of 0.0001 to 5 wt.% (e.g., 0.01 to 5 wt.%, 0.1 to 2.5 wt.%, 1 to 2 wt.%), such as no more than 2 wt.%, or no more than 1 wt.%.

[0080] In some embodiments the lipid composition disclosed herein comprises no additional excipient.Methods

[0081] The disclosure provides, in part, a method of treating a disease or condition selected from the group consisting of Alzheimer's disease, hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive disorders, heart failure, obstructive sleep apnea, osteoarthritis, chronic lower back pain, chronic kidney disease, metabolic dysfunction- associated steatotic liver disease (MASLD), the reduction of major adverse cardiac events, and / or the reduction in body weight, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia, Parkinson's disease, MASH, polycystic ovary syndrome and gastric ulcers comprising administering a lipid composition comprising from about 0.1 mg - 100 mg (e.g., about 2 mg, about 4 mg, about 6 mg, about 8 mg, about 9 mg, about 10mg, about 12 mg, about 14 mg, about 16 mg, about 18 mg, about 20 mg, about 22 mg, about 24 mg, about 26 mg, , about 28 mg, about 30 mg, about 32 mg, about 34 mg, about 38 mg, about 40 mg, about 42 mg, about 44 mg, about 46 mg, about 48 mg, about 50 mg, about 52 mg, about 54 mg, about 56 mg, about 58 mg, about 60 mg, about 62 mg, about 64 mg, about 66 mg, about 68 mg, about 70 mg) of retatrutide, or an equivalent amount of a pharmaceutically acceptable salt thereof (as an active ingredient), one or more pharmaceutically acceptable lipid excipients, and one or more solvents to a patient. In some embodiments the administration is once every three to five weeks, such as once every 28 days ± 7 days, such as once every 28 days ± 6 days, such as once every 28 days ± 5 days, such as once every 28 days ± 4 days, such as once every 28 days ± 3 days, such as once every 28 days ± 2 days, such as once every 28 days ± 1 days.

[0082] In some embodiments the administration is once every four weeks, such as once every 28 days ± 3 days, such as once every 28 days ± 2 days, such as once every 28 days ± 1 days, such as once every 28 days. In an embodiment the method is for treating diabetes, obesity, heart failure, obstructive sleep apnea, osteoarthritis, chronic lower back pain, chronic kidney disease, metabolic dysfunction-associated steatotic liver disease (MASLD), the reduction of major adverse cardiac events, and / or the reduction in body weight.

[0083] Viscosity

[0084] The lipid compositions described herein can have a viscosity suitable for subcutaneous injection. In some embodiments, the lipid compositions can have a viscosity at 25 °C of less than about 1000 mPa- s, less than about 750 mPa-s, less than about 600 mPa-s, less than about 500 mPa- s, less than about 400 mPa-s, less than about 350 mPa- s, less than about 300 mPa-s, less than about 250 mPa- s, less than about 200 mPa- s, less than about 150 mPa-s, less than about 100 mPa-s, less than about 50 mPa-s, about 100 mPa-s to about 500 mPa-s, about 200 mPa-s to about 350 mPa-s, or from about 150 mPa-s to about 450 mPa-s.

[0085] Total Solvent Amount

[0086] As described herein, the lipid composition can comprise one or more solvents, such as ethanol, DMSO, propylene glycol, etc. It has been unexpectedly found that the total amount of the solvent can impact the rate of release of the active ingredient, retatrutide. It has been unexpectedly found that the total amount of solvent can impact the Cmax and / or initial release of retatrutide from the lipid composition. It can be desirable for the lipid composition to slowly release the retatrutide to provide an extended releaseof the retatrutide. It has been unexpectedly found that low levels of total solvent allow for a slow rate of release of retatrutide and / or low level of viscosity suitable for subcutaneous injection, while maintaining adequate levels of solubility and pharmacokinetic profile needed for the use of retatrutide as a monthly therapeutic with an acceptable initial release. Low levels of solvents can include less than about 20 wt%, less than about 15 wt%, less than about 10 wt%, less than about 5 wt%, from about 5 wt% to about 20 wt%, from about 10 wt% to about 20 wt%, from about 14 wt % to about 16 wt%, or about 15 wt% by weight of the lipid composition, of the solvent. The solvent can include one or more solvents, (e.g. a combination of two solvents) as further described herein.

[0087] Improved solubility of retatrutide can be observed when retatrutide with pharmaceutical grade purity is used. Retatrutide with pharmaceutical grade purity can include retatrutide that is suitable for use as an active pharmaceutical ingredient as determined by any medical regulatory agency in a jurisdiction of interest, such as, but not limited to the United States Food and Drug Administration or the European Medical Agency. It is believed that pharmaceutical grade retatrutide can lead to improved retatrutide solubility by minimizing impurities that may be difficult to solubilize.

[0088] It has been unexpectedly found that improved solubility of retatrutide can be observed when retatrutide substantially free of impurities is used. Substantially free of impurities can mean that the retatrutide can have less than 1 wt%, less than 0.1 wt%, less than 0.01 wt %, or less than 0.001 wt %, by weight of the tirzepatide, can be used. It is believed that pharmaceutical grade retatrutide can lead to improved tirzepatide solubility by minimizing impurities that may be difficult to solubilize.

[0089] Exemplary Embodiments

[0090] El. A composition comprising about 1 wt.% to about 8 wt.% or about 2 to about 80 mg retatrutide or a pharmaceutically acceptable salt thereof, about 30 wt.% to about 55 wt.% diacyl glycerol, about 30 wt.% to about 55 wt.% of phosphatidyl choline, about 5 wt.% to about 40 wt% of a solvent or a solvent mixture.

[0091] E2. The composition of El, wherein the solvent or solvent mixture is selected from the group consisting of ethanol, propylene glycol (PG), dimethyl sulfoxide (DMSO), N-methyl-2-pyrrolidone (NMP), and glycerol.

[0092] E3. The composition of any one of E1-E2, wherein the solvent is a mixture comprising at least

[0093] a) ethanol and DMSO, such as about 2 to about 20 wt.% ethanol and about 1 to about 20 wt.% DMSO,

[0094] b) ethanol and PG, such as about 2 to about 20 wt.% ethanol and about 1 to about 20 wt.% PG, or

[0095] c) ethanol and NMP, such as about 2 to about 20 wt.% ethanol and about 1 to about 20 wt.% NMP.

[0096] E4. The composition of any one of E1-E3, wherein the solvent is a mixture of ethanol andDMSO, such as about 2 to about 20 wt.% ethanol and about 1 to about 20 wt.% DMSO.

[0097] E5. The composition of any one of E1-E3, wherein DMSO is present in an amount of about1 to about 10 wt.%.

[0098] E6. The composition of any one of E1-E3, wherein DMSO is present in an amount of about to 1 to about 7.5 wt.%.

[0099] E7. The composition of any one of E1-E3, wherein DMSO is present in an amount of about1 to about 5 wt.%.[000100] E8. The composition of any one of E1-E3, wherein DMSO is present in an amount of about1 wt.%, about 2 wt.%, about 3 wt.%, about 4 wt.%, about 5 wt.%, about 6 wt.%, about 7 wt.%, about 8 wt.%, about 9 wt.%, or about 10 wt.%.[000101] E9. The composition of any one of E1-E2, wherein the solvent is a mixture of ethanol andPG, such as 2-20 wt.% ethanol and 1-20 wt.% PG.[000102] E10. The composition of any one of E1-E2, and E9, wherein PG is present in an amount of about 3 to about 15 wt.%.[000103] El l. The composition of any one of E1-E2, and E9, wherein PG is present in an amount of about 3 to about 12.5 wt.%.[000104] E12. The composition of any one of E1-E2, and E9, wherein PG is present in an amount of about 5 to about 10 wt.%.[000105] E13. The composition of any one of E1-E2, and E9, wherein PG is present in an amount of about 2 wt.%, about 3 wt.%, about 4 wt.%, about 5 wt.%, about 6 wt.%, about 7 wt.%, about 8 wt.%, about 9 wt.%, about 10 wt.%, about 11 wt.%, about 12 wt.%, about 13 wt.%, about 14 wt.%, or about 15 wt.%.[000106] E14. The composition of any one of claims E1-E2, wherein the solvent is a mixture of ethanol and NMP, such as 2-20 wt.% ethanol and 1-20 wt.% NMP.[000107] E15. The composition of any one of E1-E2, and E14, wherein NMP is present in an amount of about 3 to about 15 wt.%.[000108] E16. The composition of any one of E1-E2, and E14, wherein NMP is present in an amount of about 3 to about 12.5 wt.%.[000109] E17. The composition of any one of E1-E2, and E14, wherein NMP is present in an amount of about 5 to about 10 wt.%.[000110] E18. The composition of any one of E1-E2, and E14, wherein NMP is present in an amount of about 2 wt.%, about 3 wt.%, about 4 wt.%, about 5 wt.%, about 6 wt.%, about 7 wt.%, about 8 wt.%, about 9 wt.%, about 10 wt.%, about 11 wt.%, about 12 wt.%, about 13 wt.%, about 14 wt.%, or about 15 wt.%.[000111] E19. The composition of any one of E1-E18, wherein ethanol is present in an amount of about 1 to about 20 wt.%.[000112] E20. The composition of any one of E1-E19, wherein ethanol is present in an amount of about 3 to about 15 wt.%.[000113] E21. The composition of any one of E1-E19, wherein ethanol is present in an amount of about 3 to about 12.5 wt.%.[000114] E22. The composition of any one of E1-E19, wherein ethanol is present in an amount of about 5 to about 10 wt.%.[000115] E23. The composition of any one of E1-E19, wherein ethanol is present in an amount of about 2 wt.%, about 3 wt.%, about 4 wt.%, about 5 wt.%, about 6 wt.%, about 7 wt.%, about 8 wt.%, about 9 wt.%, about 10 wt.%, about 11 wt.%, about 12 wt.%, about 13 wt.%, about 14 wt.%, or about 15 wt.%.[000116] E24. The composition of any one of E1-E23 comprising about 2 to about 7 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.[000117] E25. The composition of any one of El-23 comprising about 2 to about 6 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.[000118] E26. The composition of any one of E1-E23 comprising about 2 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.[000119] E27. The composition of any one of E1-E23 comprising about 3 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.[000120] E28. The composition of any one of E1-E23 comprising about 4 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.[000121] E29. The composition of any one of E1-E23 comprising about 5 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.[000122] E30. The composition of any one of E1-E23 comprising about 6 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.[000123] E31. The composition of any one of E1-E23 comprising about 7 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.[000124] E32. The composition of any one of E1-E23 comprising about 4 mg to about 70 mg of retatrutide or a pharmaceutically acceptable salt thereof.[000125] E33. The composition of any one of E1-E23 comprising about 8 mg to about 60 mg of retatrutide or a pharmaceutically acceptable salt thereof.[000126] E34. The composition of any one of E1-E23 comprising about 12 mg to about 48 mg of retatrutide or a pharmaceutically acceptable salt thereof.[000127] E35. The composition of any one of E1-E23 comprising about 2 mg, about 4 mg, about 6 mg, about 8 mg, about 9 mg, about 10 mg, about 12 mg, about 14 mg, about 16 mg, about 18 mg, about 20 mg, about 22 mg, about 24 mg, about 26 mg, about 28 mg, about 30 mg, about 32 mg, about 34 mg, about 36 mg, about 40 mg, about 42 mg, about 44 mg, about 46 mg, about 48 mg, about 50 mg, about 52 mg, about 54 mg, about 56 mg, about 58 mg, or about 60 mg of retatrutide or a pharmaceutically acceptable salt thereof[000128] E36. The composition of any one of E1-E35, wherein the composition is formulated for subcutaneous injection.[000129] E37. The composition of any one of E1-E35, for use as a medicament.[000130] E38. The composition of any one of E1-E35, for use as an extended-release medicament providing a release of retatrutide or a pharmaceutically acceptable salt thereof for about at least one to six weeks, such as about two to six weeks, such as about two to five weeks, such as about three to five weeks, such as once every 28 days ± 7 days, such as once every 28 days ± 6 days, such as once every 28 days ± 5 days, such as once every 28 days ± 4 days, such as once every 28 days ± 3 days, such as once every 28 days ± 2 days, such as once every 28 days ± 1 days, such as once every 28 days provides a therapeutic dose of Retatrutide over at least 28 days upon human administration.[000131] E39. The composition of any one of E1-E35, for use as an extended-release medicament providing a release of retatrutide or a pharmaceutically acceptable salt thereof for about 28 days ± 7 days. [000132] E40. The composition of any one of 1-35, wherein the composition is formulated for subcutaneous injection providing an extended release of retatrutide or a pharmaceutically acceptable salt thereof for about 28 days to about 33 days after administered to a human.[000133] E41. The composition of any one of E1-E35, for treating a disease or condition selected from the group consisting of Alzheimer's disease, hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive disorders, heart failure, obstructive sleep apnea, osteoarthritis, chronic lower back pain, chronic kidney disease, metabolic dysfunction-associated steatotic liver disease (MASLD), the reduction of major adverse cardiac events, the reduction in body weight, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia, Parkinson's disease, polycystic ovary syndrome, MASH (metabolic dysfunction-associated steatohepatitis) and gastric ulcers. [000134] E42. A method of treating a disease or condition comprising administering, to a mammalian patient in need thereof a composition according to any one of E1-E35.[000135] E43. The method of E42, wherein the composition is administered to the mammalian patient by injection.[000136] E44. The method of E42 or 43 wherein the composition is administered to the mammalian patient by subcutaneous injection.[000137] E45. The method of any one of E41-44, wherein the composition is administered to the mammalian patient by subcutaneous injection once every one to six weeks, such as about two to six weeks, such as about two to five weeks, such as about three to five weeks, such as once every 28 days ± 7 days, such as once every 28 days ± 6 days, such as once every 28 days ± 5 days, such as once every 28 days ± 4 days, such as once every 28 days ± 3 days, such as once every 28 days ± 2 days, such as once every 28 days ± 1 days such as once every 28 days.[000138] E46. The method of any one of E41-44, wherein the composition is administered to the mammalian patient by subcutaneous injection once every 28 days ± 7 days.[000139] E47. The method of any one of E41-E46, wherein the disease or condition is selected from the group consisting of Alzheimer's disease, hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive disorders, heart failure, obstructive sleep apnea, osteoarthritis, chronic lower back pain, chronic kidney disease, metabolicdysfunction-associated steatotic liver disease (MASLD), the reduction of major adverse cardiac events, the reduction in body weight, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia, Parkinson's disease, polycystic ovary syndrome, MASH, and gastric ulcers.[000140] E48. A method of treating a mammalian patient suffering from a disease or condition wherein an first dose of the composition of any one of El -35 is administered to the mammalian patient, and wherein a second or subsequent dose of the composition of any one of claims 1-35 is administered after about 1-7 days, or 1 to 4 weeks, such as 1, 2, 3 or 4 weeks after the administration of the first dose, and wherein the amount of the second dose is the same or higher than the amount of the first dose.[000141] E49. The method according to E48, wherein the method is an initiation method, which is administered one or more times to the mammalian patient, and wherein the method is repeated until a maintenance dose of the composition of any one of claims 1-35 is administered, and wherein the maintenance dose is administered after about three to five weeks, such as once every 28 days ± 7 days, such as once every 28 days ± 6 days, such as once every 28 days ± 5 days, such as once every 28 days ± 4 days, such as once every 28 days ± 3 days, such as once every 28 days ± 2 days, such as once every 28 days ± 1 days such as once every 28 days after the first, second or subsequent dose and wherein the amount of the maintenance dose is the same or higher than the amount of the first, second or subsequent dose.[000142] E50. The composition of any one of E1-E35, wherein the viscosity is about 50 - about 450 mPa s.[000143] E51. The composition of any one of E1-E35, wherein the lipid composition is a solution, such as a clear solution as determined by visual inspection.[000144] E52. The composition of any one of E1-E35, wherein the composition is provided in a dose volume of about 0.1 - about 1.5 mL, such as about 1.0 mL, such as 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL, 0.6 mL, 0.7 mL, 0.8 mL, 0.9 mL, 1.0 mL, 1.1 mL, 1.2 mL, 1.3 mL, 1.4 mL, or 1.5 mL.[000145] E53. The method according to any one of E48 and 49, wherein the maintenance dose is administered about once every month, for example once every 28 days ± 7 days, such as once every 28 days ± 6 days, such as once every 28 days ± 5 days, such as once every 28 days ± 4 days, such as once every 28 days ± 3 days, such as once every 28 days ± 2 days, such as once every 28 days ± 1 days such as once every 28 days.EXAMPLE 1FORMULATION EXAMPLE[000146] Materials[000147] RET Retatrutide[000148] SPC Soy phosphatidylcholine[000149] GDO Glycerol dioleate[000150] EtOH Ethanol[000151] PG Propylene glycol[000152] DMSO Dimethyl sulfoxide[000153] Preparation of formulations[000154] Lipid placebo mixtures were prepared by weighing appropriate amounts of SPC, GDO and solvents into sterilized glass vials. Sealed vials were then placed on a roller mixer by end-over-end rotation at room temperature (RT) until mixed completely into clear homogeneous liquid solutions (<24 hours). GIP powder was then added to the respective lipid placebo formulations in new glass vials. Vials were sealed and placed on mixing at RT for 2-24 hours.[000155] Viscosity Measurements[000156] Viscosity measurements were performed on a Brookfield CAP2000+ viscometer instrument. All the measurements were performed at 25 °C with 10 s hold time and 10 s run time at shear rates of 1333 to 4000 s’1. Sample volume used was 100 pL.[000157] Evaluation of RET solubility in lipid compositions[000158] The solubility was assessed by adding RET peptide powder to respective lipid placebo mixtures followed by mixing at RT for 2-24 hours. After preparation, the solubility of RET was assessed by visual inspection in Tables 1-9. Results showed that RET has good solubility in the lipid compositions and that a drug load of at least 8 wt.% is feasible.Table 1. Solubility of RET in various lipid compositions (wt.%)Table 1: Solubility evaluation of RET in FluidCrystal formulations of different compositions (all components in wt%)Table 2: Solubility evaluation of RET in lipid formulations of different compositions (all components in wt%)RET = retatrutide; SPC = soy phosphatidylcholine; GDO = glycerol dioleate; EtOH = ethanol; PG = propylene glycol; DMSO = dimethyl sulfoxide; NMP = N-methyl pyrrolidone; PEG 300 = poly(ethylene glycol) 300; BzOH = benzyl alcohol; BzBz = benzyl benzoate; * piece of API stuck on vialEXAMPLE 2IN VITRO RELEASE (IVR)[000159] In vitro release experiments were performed in vials by injecting 200 pL of each of the formulations into 10 mL of PBS and equilibrating the depots using very gentle orbital movement for up to a week at 37°C. Determination of RET in PBS was carried out by gradient HPLC with UV detection.[000160] Table 13. In vitro release data of RET (50 mg / ml) into PBS in vials. Results are shown as % of RET released into PBS (average values of three replicates +SE).[000161] Table4. In vitro release data of RET (65 mg / ml) into PBS in vials. Results are shown as % of RET released into PBS (average values of three replicates +SE).EXAMPLE 3IN VIVO PHARMACOKINETICS[000163] The pharmacokinetics (PK) of RET after single dose administration of one formulation sample (composition S03, Table 1) to rats were evaluated.[000164] The formulation was prepared by initially mixing all the excipients (i.e., lipids and solvents) except the peptide until a homogeneous solution was obtained.[000165] The necessary amount of peptide powder was weighed into a separate vial, followed by mixing with the lipid solution prepared as described above. The content was mixed until a new homogeneous solution was generated.[000166] The RET-containing formulation obtained was sterile-filtered through 0.22 pm pore filter into a sterile vial.[000167] The RET-containing formulation was administered subcutaneously to rats (dose: 1 mg / rat; dose volume: 0.1 mL / rat).[000168] Blood samples were collected from the rats dosed with the RET-containing formulation at different time points after administration. Plasma was separated by standard procedures and stored at - 80°C until analysis.[000169] Determination of RET content in plasma samples was performed by a liquid chromatography (LC) - mass spectrometry (MS) method, qualified for determination of RET in rat plasma (Lower Limit of Quantification (LLOQ) was 5 ng / mL).The average (n=3) PK profile of RET after a single dose administration of the composition S03 (Table1) to rats is disclosed in Fig. 1 and Table 10.Table 10. PK Profile of RETINCORPORATION BY REFERENCE[000170] All publications and patents mentioned herein, including those items listed below, are hereby incorporated by reference in their entirety for all purposes as if each individual publication or patent was specifically and individually incorporated by reference. In case of conflict, the present application, including any definitions herein, will control.EQUIVALENTS[000171] While specific embodiments of the subject disclosure have been discussed, the above specification is illustrative and not restrictive. Many variations of the disclosure will become apparent to those skilled in the art upon review of this specification. The full scope of the disclosure should be determined by reference to the claims, along with their full scope of equivalents, and the specification, along with such variations.[000172] Unless otherwise indicated, all numbers expressing quantities of ingredients, reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth in this specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained by the present disclosure.

Claims

-32-WHAT IS CLAIMED IS:

1. A composition comprising about 1 wt.% to about 8 wt.% or about 2 to about 80 mg retatrutide or a pharmaceutically acceptable salt thereof, about 30 wt.% to about 55 wt.% diacyl glycerol, about 30 wt.% to about 55 wt.% of phosphatidyl choline, about 5 wt.% to about 40 wt% of a solvent or a solvent mixture.

2. The composition of claim 1, wherein the solvent or solvent mixture is selected from the group consisting of ethanol, propylene glycol (PG), dimethyl sulfoxide (DMSO), N-methyl-2- pyrrolidone (NMP), and glycerol.

3. The composition of any one of claims 1-2, wherein the solvent is a mixture comprising at least a) ethanol and DMSO, such as about 2 to about 20 wt.% ethanol and about 1 to about 20 wt.% DMSO, b) ethanol and PG, such as about 2 to about 20 wt.% ethanol and about 1 to about 20 wt.% PG, or c) ethanol and NMP, such as about 2 to about 20 wt.% ethanol and about 1 to about 20 wt.% NMP.

4. The composition of any one of claims 1-2, wherein the solvent is a mixture of ethanol and DMSO, such as about 2 to about 20 wt.% ethanol and about 1 to about 20 wt.% DMSO.

5. The composition of any one of claims 1-3, wherein DMSO is present in an amount of about 1 to about 10 wt.%.

6. The composition of any one of claims 1-3, wherein DMSO is present in an amount of about to 1 to about 7.5 wt.%.

7. The composition of any one of claims 1-3, wherein DMSO is present in an amount of about 1 to about 5 wt.%.

8. The composition of any one of claims 1-3, wherein DMSO is present in an amount of about 1 wt.%, about 2 wt.%, about 3 wt.%, about 4 wt.%, about 5 wt.%, about 6 wt.%, about 7 wt.%, about 8 wt.%, about 9 wt.%, or about 10 wt.%.-33-9. The composition of any one of claims 1-2, wherein the solvent is a mixture of ethanol and PG, such as 2-20 wt.% ethanol and 1-20 wt.% PG.

10. The composition of any one of claims 1-2, and 9, wherein PG is present in an amount of about 3 to about 15 wt.%.

11. The composition of any one of claims 1-2, and 9, wherein PG is present in an amount of about 3 to about 12.5 wt.%.

12. The composition of any one of claims 1-2, and 9, wherein PG is present in an amount of about 5 to about 10 wt.%.

13. The composition of any one of claims 1-2, and 9, wherein PG is present in an amount of about 2 wt.%, about 3 wt.%, about 4 wt.%, about 5 wt.%, about 6 wt.%, about 7 wt.%, about 8 wt.%, about 9 wt.%, about 10 wt.%, about 11 wt.%, about 12 wt.%, about 13 wt.%, about 14 wt.%, or about 15 wt.%.

14. The composition of any one of claims 1-2, wherein the solvent is a mixture of ethanol and NMP, such as 2-20 wt.% ethanol and 1-20 wt.% NMP.

15. The composition of any one of claims 1-2, and 14, wherein NMP is present in an amount of about 3 to about 15 wt.%.

16. The composition of any one of claims 1-2, and 14, wherein NMP is present in an amount of about 3 to about 12.5 wt.%.

17. The composition of any one of claims 1-2, and 14, wherein NMP is present in an amount of about 5 to about 10 wt.%.

18. The composition of any one of claims 1-2, and 14, wherein NMP is present in an amount of about 2 wt.%, about 3 wt.%, about 4 wt.%, about 5 wt.%, about 6 wt.%, about 7 wt.%, about 8 wt.%, about 9 wt.%, about 10 wt.%, about 11 wt.%, about 12 wt.%, about 13 wt.%, about 14 wt.%, or about 15 wt.%.

19. The composition of any one of claims 1-18, wherein ethanol is present in an amount of about 1 to about 20 wt.%.

20. The composition of any one of claims 1-19, wherein ethanol is present in an amount of about 3 to about 15 wt.%.

21. The composition of any one of claims 1-19, wherein ethanol is present in an amount of about 3 to about 12.5 wt.%.

22. The composition of any one of claims 1-19, wherein ethanol is present in an amount of about 5 to about 10 wt.%.

23. The composition of any one of claims 1-19, wherein ethanol is present in an amount of about 2 wt.%, about 3 wt.%, about 4 wt.%, about 5 wt.%, about 6 wt.%, about 7 wt.%, about 8 wt.%, about 9 wt.%, about 10 wt.%, about 11 wt.%, about 12 wt.%, about 13 wt.%, about 14 wt.%, or about 15 wt.%.

24. The composition of any one of claims 1-23 comprising about 2 to about 7 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.

25. The composition of any one of claims 1-23 comprising about 2 to about 6 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.

26. The composition of any one of claims 1-23 comprising about 2 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.

27. The composition of any one of claims 1-23 comprising about 3 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.

28. The composition of any one of claims 1-23 comprising about 4 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.

29. The composition of any one of claims 1-23 comprising about 5 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.

30. The composition of any one of claims 1-23 comprising about 6 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.

31. The composition of any one of claims 1-23 comprising about 7 wt.% of retatrutide or a pharmaceutically acceptable salt thereof.

32. The composition of any one of claims 1-23 comprising about 4 mg to about 70 mg of retatrutide or a pharmaceutically acceptable salt thereof.

33. The composition of any one of claims 1-23 comprising about 8 mg to about 60 mg of retatrutide or a pharmaceutically acceptable salt thereof.

34. The composition of any one of claims 1-23 comprising about 12 mg to about 48 mg of retatrutide or a pharmaceutically acceptable salt thereof.

35. The composition of any one of claims 1-23 comprising about 2 mg, about 4 mg, about 6 mg, about 8 mg, about 9 mg, about 10 mg, about 12 mg, about 14 mg, about 16 mg, about 18 mg, about20 mg, about 22 mg, about 24 mg, about 26 mg, about 28 mg, about 30 mg, about 32 mg, about34 mg, about 36 mg, about 40 mg, about 42 mg, about 44 mg, about 46 mg, about 48 mg, about50 mg, about 52 mg, about 54 mg, about 56 mg, about 58 mg, or about 60 mg of retatrutide or a pharmaceutically acceptable salt thereof36. The composition of any one of claims 1-35, wherein the composition is formulated for subcutaneous injection.

37. The composition of any one of claims 1-35, for use as a medicament.

38. The composition of any one of claims 1-35, for use as an extended-release medicament providing a release of retatrutide or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable salt thereof for about at least one to six weeks, such as about two to six weeks, such as about two to five weeks, such as about three to five weeks, such as once every 28 days ± 7 days, such as once every 28 days ± 6 days, such as once every 28 days ± 5 days, such as once every 28 days ± 4 days, such as once every 28 days ± 3 days, such as once every 28 days ± 2 days, such as once every 28 days ± 1 days, such as once every 28 days provides a therapeutic dose of retatrutide over at least 28 days upon human administration.

39. The composition of any one of claims 1-35, wherein the composition is formulated for subcutaneous injection providing an extended release of retatrutide or a pharmaceutically acceptable salt thereof for about 28 days to about 33 days after administered to a human.-36-40. The composition of any one of claims 1-35, for treating a disease or condition selected from the group consisting of Alzheimer's disease, hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive disorders, heart failure, obstructive sleep apnea, osteoarthritis, chronic lower back pain, chronic kidney disease, metabolic dysfunction-associated steatotic liver disease (MASLD), the reduction of major adverse cardiac events, the reduction in body weight, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia, Parkinson's disease, polycystic ovary syndrome, MASH (metabolic dysfunction-associated steatohepatitis) and gastric ulcers.

41. A method of treating a disease or condition comprising administering, to a mammalian patient in need thereof a composition according to any one of claims 1-35.

42. The method of claim 41 , wherein the composition is administered to the mammalian patient by injection.

43. The method of claims 41 or 42 wherein the composition is administered to the mammalian patient by subcutaneous injection.

44. The method of any one of claims 41-43, wherein the composition is administered to the mammalian patient by subcutaneous injection once every one to six weeks, such as about two to six weeks, such as about two to five weeks, such as about three to five weeks, such as once every 28 days ± 7 days, such as once every 28 days ± 6 days, such as once every 28 days ± 5 days, such as once every 28 days ± 4 days, such as once every 28 days ± 3 days, such as once every 28 days ± 2 days, such as once every 28 days ± 1 days such as once every 28 days.

45. The method of any one of claims 41-43, wherein the composition is administered to the mammalian patient by subcutaneous injection once every 28 days ± 7 days.

46. The method of any one of claims 41-45, wherein the disease or condition is selected from the group consisting of Alzheimer's disease, hyperglycemia, type 2 diabetes, impaired glucose tolerance, type 1 diabetes, obesity, hypertension, syndrome X, dyslipidemia, cognitive disorders, heart failure, obstructive sleep apnea, osteoarthritis, chronic lower back pain, chronic kidney disease, metabolic dysfunction-associated steatotic liver disease (MASLD), the reduction of major-37- adverse cardiac events, the reduction in body weight, atherosclerosis, myocardial infarction, coronary heart disease and other cardiovascular disorders, stroke, inflammatory bowel syndrome, dyspepsia, Parkinson's disease, polycystic ovary syndrome, MASH, and gastric ulcers.

47. A method of treating a mammalian patient suffering from a disease or condition wherein an first dose of the composition of any one of claims 1-35 is administered to the mammalian patient, and wherein a second or subsequent dose of the composition of any one of claims 1-35 is administered after about 1-7 days, or 1 to 4 weeks, such as 1, 2, 3 or 4 weeks after the administration of the first dose, and wherein the amount of the second dose is the same or higher than the amount of the first dose.

48. The method according to claim 47, wherein the method is an initiation method, which is administered one or more times to the mammalian patient, and wherein the method is repeated until a maintenance dose of the composition of any one of claims 1-35 is administered, and wherein the maintenance dose is administered after about three to five weeks, such as once every 28 days ± 7 days, such as once every 28 days ± 6 days, such as once every 28 days ± 5 days, such as once every 28 days ± 4 days, such as once every 28 days ± 3 days, such as once every 28 days ± 2 days, such as once every 28 days ± 1 days such as once every 28 days after the first, second or subsequent dose and wherein the amount of the maintenance dose is the same or higher than the amount of the first, second or subsequent dose.49 The composition of any one of claims 1-35, wherein the viscosity is about 50 - about 450 mPa s.

50. The composition of any one of claims 1-35, wherein the composition is a solution, such as a clear solution as determined by visual inspection.

51. The composition of any one of claims 1-35, wherein the composition is provided in a dose volume of about 0.1 - about 1.5 mL, such as about 1.0 mL, such as 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL, 0.6 mL, 0.7 mL, 0.8 mL, 0.9 mL, 1.0 mL, 1.1 mL, 1.2 mL, 1.3 mL, 1.4 mL, or 1.5 mL.

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