Compositions containing mtor inhibitors and lipids

Abstract

The invention relates to compositions of mTOR inhibitors and administering compositions of mTOR inhibitors. Embodiments provide compositions comprising mTOR inhibitors with at least one lipid and / or guggulsterol, and / or guggulsterol derivatives, and administering the compositions in a subject.

Description

[0001] DESCRIPTION

[0002] TITLE OF THE INVENTION:COMPOSITIONS CONTAINING mTOR

[0003] INHIBITORS AND LIPIDS

[0004] FIELD OF THE INVENTION

[0005] The invention relates to compositions comprising mTOR inhibitors and lipids. The invention further relates to compositions comprising mTOR inhibitors, guggulsterol and / or guggulsterol derivatives. In some embodiments, the invention relates to composition comprising mTOR inhibitors, lipids including phosphatidylcholine, phosphatidylglycerol. In preferred embodiments, the invention relates to compositions comprising mTOR inhibitors, phosphatidylcholine, and guggulsterol or guggulsterol derivatives. The invention further relates administering composition to a human subject in the treatment or prevention of diseases. The invention is further related to administering the composition to human subject to achieve comparable pharmacokinetic profile compared to the solvent-based commercially available temsirolimus injection product. Compositions according to the present invention are suitable for practice on an industrial manufacturing scale, and may be practiced, e.g., as a continuous method.

[0006] BACKGROUND OF THE INVENTION

[0007] The mammalian target of rapamycin (mTOR) is a member of the phosphatidylinositol 3 -kinase-related kinase (PIKK) family, which is one of the key players of cellular metabolism that is coupled with nutrient availability, energy, and homeostasis. It plays an important role in cell growth, differentiation, metastasis, and survival, and has become an important target of cancer treatment. Several therapeutic agents have been developed which inhibit mTOR and commonly referred to as mTOR inhibitors including Rapamycin and its analogs. Rapamycin, also known as sirolimus, a macrolide lactone, was initially described as an antifungal agent. However, it also exhibits immunosuppressant, cytostatic, antiangiogenic, and antiproliferative properties, expanding the clinical applications to organ transplantation and oncological fields. Sirolimus is initially approved for oral administration as immunosuppressant and indicated for the prophylaxis of organ rejection in patients aged >13 years receiving renal transplants. It is also used to coat coronary stents and to treat a rare lung disease called Lymphangioleiomyomatosis (LAM). Recently, sirolimus, protein-bound particles developed for intravenous use, has been approved for the treatment of adult patients with locally advanced unresectable or metastatic malignant perivascular epithelioid cell tumor (PEComa). Most recently, it is also approved for topical treatment in the form of 0.2% topical gel for facial angiofibroma associated with tuberous sclerosis.

[0008] To explore the therapeutic profile, the most common rapamycin analogs developed include everolimus, temsirolimus, deforolimus, and zotarolimus. While deforolimus, and zotarolimus remains as investigational candidates, everolimus is approved to treat postmenopausal advanced hormone receptor-positive, HER2-negative breast cancer in women, progressive neuroendocrine tumors of pancreatic origin (PNET), advanced renal cell carcinoma (RCC) after failure of treatment with sunitinib or sorafenib, renal angiomyolipoma (AML), tuberous sclerosis complex (TSC), and subependymal giant cell astrocytoma (SEGA) associated with TSC as well as renal and liver transplantation. Temsirolimus is approved as a first-line treatment for advanced renal cell cancer. Sirolimus, an active metabolite of temsirolimus, is the principal metabolite in humans following intravenous treatment.

[0009] Tacrolimus is also a macrolide lactone structurally similar to sirolimus and is indicated for the prophylaxis of organ rejection in adult and pediatric patients receiving allogeneic liver, kidney, heart, or lung transplants, in combination with other immunosuppressants. It is also used as an ointment in the treatment of eczema, in particular atopic dermatitis. Because of the structural similarity between tacrolimus and sirolimus, and the ability of both to bind to FK506 binding protein 12 (FKBP12), it is hypothesized that tacrolimus can also inhibit the mTOR signaling, constituting a possible mechanism of 0 cell toxicity.

[0010] Temsirolimus is marketed as Torisel® and is approved for the treatment of advanced renal cell carcinoma. The recommended dose of TORISEL for advanced renal cell carcinoma is 25 mg administered as an intravenous infusion over a 30 - 60-minute period once a week.

[0011] Torisel® is supplied as a sterile non-aqueous clear solution (concentrate) for injection containing Temsirolimus 25 mg / mL, in dehydrated alcohol (39.5% w / v) and propylene glycol (50.3% w / v). The product is provided with an additional vial as diluent containing polysorbate 80 (40.0% w / v), polyethylene glycol 400 (PEG400) (42.8% w / v), and dehydrated alcohol (19.9% w / v). The concentrate and the diluent are intended for the preparation of a premix solution of temsirolimus at 10 mg / ml prior to dilution with 250 mL of 0.9% saline solution in an infusion bag. The generic versions of Torisel® are also approved for marketing.

[0012] Temsirolimus is highly lipophilic and practically insoluble in water. Due to its insolubility, various solubilizers such as polysorbate 80, PEG400, propylene glycol, and dehydrated alcohol were successfully exploited to formulate for intravenous administration. However, hypersensitivity reactions and infusion related toxicities are associated with the use of Polysorbate 80, propylene glycol, and ethanol, and to reduce the risk of these side effects, patients are routinely premedicated with prior to the treatment of temsirolimus.

[0013] To circumvent the toxic effect of mTOR inhibitors and the excipients in the current marketed intravenous product, new compositions of mTOR inhibitors such as temsirolimus for parenteral route of administration is warranted.

[0014] SUMMARY OF THE INVENTION

[0015] This invention provides compositions containing mTOR inhibitors. In some embodiments, the composition comprises mTOR inhibitor, phosphatidylcholine and or phosphatidylglycerol. In some embodiments, the composition further comprises mTOR inhibitor and one or more of guggulsterol, guggulsterol derivatives. In some embodiments, the composition comprises mTOR inhibitor, phosphatidylcholine and or phosphatidylglycerol, and one or more of guggulsterol, guggulsterol derivatives. In some preferred embodiments, the composition further comprises other excipients. Certain embodiments comprise a composition comprising mTOR inhibitors and administering the composition to a subject. In certain embodiments, the subject is a mammal. In preferred embodiments, the subject is human.

[0016] In some embodiments, this invention provides compositions comprising mTOR inhibitor and phosphatidylcholine for parenteral delivery to a human subject. In some embodiments, the composition for parenteral delivery is in the form of suspension. In certain embodiments compositions of the present invention comprise mTOR inhibitor and phosphatidylglycerol for parenteral delivery to a human subject. In some embodiments, the composition for parenteral delivery is in the form of suspension.

[0017] Provided herein in some embodiments are compositions comprising mTOR inhibitor and guggulsterol for parenteral delivery to a human subject. In some embodiments, the composition for parenteral delivery is in the form of suspension.

[0018] The invention in some embodiments provides compositions comprising mTOR inhibitor and a guggulsterol derivative for parenteral delivery to a human subject. In some embodiments, parenteral delivery is in the form of suspension.

[0019] The invention in some embodiments provides compositions comprising mTOR inhibitor, phosphatidylcholine, and guggulsterol for parenteral delivery to a human subject. In some embodiments, the parenteral delivery is in the form of suspension.

[0020] The invention in some embodiments provides compositions comprising mTOR inhibitor, phosphatidylcholine, and guggulsterol derivative for parenteral delivery to a human subject. In some embodiments, the parenteral delivery is in the form of suspension.

[0021] The invention in some embodiments provides compositions comprising mTOR inhibitor, phosphatidylglycerol, and guggulsterol for parenteral delivery to a human subject. In some embodiments, the parenteral delivery is in the form of suspension.

[0022] The invention in some embodiments provides compositions comprising mTOR inhibitor, phosphatidylglycerol, and guggulsterol derivative for parenteral delivery to a human subject. In some embodiments, the parenteral delivery is in the form of suspension.

[0023] In some embodiments, this invention provides compositions comprising mTOR inhibitor, phosphatidylcholine, and phosphatidylglycerol for parenteral delivery to a human subject. In some embodiments, the composition for parenteral delivery is in the form of suspension.

[0024] In some embodiments, this invention provides compositions comprising mTOR inhibitor, phosphatidylcholine, phosphatidylglycerol, and guggulsterol for parenteral delivery to a human subject. In some embodiments, the composition for parenteral delivery is in the form of suspension.

[0025] In some embodiments, this invention provides compositions comprising mTOR inhibitor, phosphatidylcholine, phosphatidylglycerol, and guggulsterol derivative for parenteral delivery to a human subject. In some embodiments, the composition for parenteral delivery is in the form of suspension.

[0026] In some embodiments, this invention provides compositions comprising mTOR inhibitor, and lipid(s) for rectal delivery to a human subject. In some embodiments, the composition for rectal delivery is in the form of suspension.

[0027] In some embodiments, a mTOR inhibitors composition of present invention is coadministered with other drugs. Drugs that can be co-administered along with mTOR inhibitors composition include but are not limited to anticancer drugs such as doxorubicin, epirubicin, methotrexate, mitoxantrone, capecitabine, carboplatin, cisplatin, etoposide, 5-flurouracil, cyclophosphamide, daunomycin, bleomycin, gemcitabine, irinotecan, SN-38, mitoxantrone, cytrabine, capecitabine, mitomycin, sunitinib, sorafenib, tivozanib, ibrutinib, imatinib, erlotinib, acalabrutinib, carbozantinib, bevacizumab, paclitaxel, docetaxel, cabazitaxel, vincristine, abiraterone, bicalutamide, flutamide, etc.; antihypertensive agents, such as dihydropyridines, antidepressants, antiallergic agents, etc.; corticosteroids drugs such as prednisone, methylprednisolone, dexamethasone, budesonide, hydrocortisone, etc.; antihistamine drugs such as diphenhydramine, chlorpheniramine, dexchlorpheniramine, cetirizine, levocetrizine, loratadine, desloratadine, etc. drugs for treating acid reflux such as cimetidine, ranitidine, famotidine, esomeprazole, lansoprazole, omeprazole, pantoprazole, rabeprazole, etc.

[0028] The amount of mTOR inhibitor included in a mTOR inhibitors composition according to present invention is not limited to any amount or percentage (by weight) of the final composition or weight. In some embodiments, the proportion of mTOR inhibitor is about 0.1% to about 90% of the total weight, preferably about 0.5% to about 75% of the total weight, more preferably about 1% to about 50% of the total weight. The amount of lipid(s) included in a mTOR inhibitor composition according to present invention is not limited to any particular amount or percentage (by weight) of the final composition or weight. In some embodiments, the proportion of total lipid is about 4% to about 100% by weight of total lipid, preferably about 5% to about 60% by weight of total lipid or more preferably about 8% to about 50% by weight of total lipid.

[0029] DEFINITIONS

[0030] To facilitate an understanding of the present invention, a few terms and phrases are defined below:

[0031] As used herein, the term “composition” “preparation” or “formulation” refers to the combination of an active agent (e.g, an active pharmaceutical compound) with a carrier, inert or active, excipients, making the composition especially suitable for diagnostic or therapeutic use in vitro, in vivo, or ex vivo.

[0032] As used herein, the term “active” as used in reference to an agent, composition, or compound, refers to an agent that, upon administration or application, causes a beneficial, desired, or expected result. The administration may be in one or more administrations, applications, dosages and is not intended to be limited to a particular formulation or administration route. The term is not limited to any level of activity. For example, a formulation of an active agent need not have the same level of activity as a different formulation of an active agent, so long as the active agent in the formulation is sufficiently active that an effective amount of the active agent can be administered by administration of the formulation of the agent.

[0033] The terms “agent” and “compound” are used herein interchangeably to refer to any atom, molecule mixture or more complex composition having an attributed feature. For example, an ‘active agent” or “active compound” refers to any atom, molecule, preparation mixture, etc. that, upon administration or application, causes beneficial, desired, or expected result.

[0034] The terms "pharmaceutically acceptable" or "pharmacologically acceptable," as used herein, refer to compositions that do not substantially produce adverse reactions, e.g, toxic, allergic, or immunological reactions, when administered to a subject. As used herein, the term “administration” or “administering” refers to the act of giving a drug, or active agent, or therapeutic treatment (e.g., composition of the present invention) to a physiological system (e.g., a subject or in vivo, in vitro, or ex vivo cells, tissues, and organs). Exemplary routes of administration to the human body can be through mouth (oral), skin (transdermal), eyes (ophthalmic), nose (nasal), and the like. Administration may be in one or more administrations, applications, or dosages, and is not intended to be limited to a particular administration route.

[0035] As used herein, the term “co-administration” refers to the administration of at least two agents(s) (e.g., two separate compositions, containing different active agents) or therapies to a subject. In some embodiments, the co-administration of two or more agents or therapies are concurrent. In other embodiments, a first agent / therapy is administered prior to a second agent / therapy. Those of skill in the art understand that the formulations and / or routes of administration of the various agents or therapies used may vary. The appropriate dosage for co-administration can be readily determined by one skilled in the art.

[0036] As used herein, the term “parenteral” refers to non-oral means of administration. The common parenteral route of administration includes intravenous (IV), intramuscular (IM), and subcutaneous (SC).

[0037] As used herein, the term “disease” refers to a state, signs, and / or symptoms that are associated with any impairment of the normal state of a living animal or any of its organs or tissues that interrupts or modifies the performance of normal functions and may be a response to environmental factors.

[0038] As used herein, the term “treatment” or grammatical equivalents encompasses the improvement and / or reversal of the symptoms of disease (e.g., cancer), or reduction of risk of occurrence of disease. A compound which causes an improvement in any parameter associated with disease when used in the screening methods of the instant invention may thereby be identified as a therapeutic compound. The term “treatment” refers to therapeutic treatment. For example, those who may benefit from treatment with compositions of the present invention include those already with a disease and / or disorder (e.g., cancer, or symptoms or pathologies consistent with cancer). As used herein, the term “pharmacokinetic” refers to the passage of active agents or drugs after administration into body, through it, and out of body. The passage include absorption, for example, how the active agent or drug moves from the site of administration to the site of action; distribution, for example, the journey of the active agent or drug through the bloodstream to various tissues of the body; metabolism, for example, the process that breaks down the drug inside body; and excretion, for example, the removal of the drug from the body.

[0039] The use of terms “a” and “an” and “the” and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms “comprising”, “including”, “having”, and “containing” are to be construed as open-ended terms (i.e., meaning “including but limited to”) unless otherwise noted. The use of any and all examples, or exemplary language (e.g., ‘such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specifications should be constructed as indicating any non-claimed element as essential to the practice of the invention.

[0040] BRIEF DESCRIPTION OF DRAWINGS

[0041] [FIG. 1 ] FIG. 1 is a graph showing the blood concentrations of temsirolimus over time in Example 13.

[0042] [FIG. 2] FIG. 2 is a graph showing the blood concentrations of sirolimus over time in Example 13.

[0043] DETAILED DESCRIPTION OF THE INVENTION

[0044] The invention relates to a composition comprising mTOR inhibitors formulation. In some embodiment, the invention comprises administrating a mTOR composition to a human subject, e.g., to treat a disease. In some embodiments, the composition comprising mTOR inhibitors comprises lipids, for example, phosphatidylcholine or phosphatidylglycerol. In certain embodiments, the composition comprising mTOR inhibitors comprises guggulsterol or a guggulsterol derivative. In other embodiments, the composition comprises phosphatidylcholine or phosphatidylglycerol and / or guggulsterol, a guggulsterol derivative.

[0045] Embodiments of the invention are described in the Summary, and in this Detailed Description of the Invention. Although the invention has been described in connection with specific embodiments, the invention as claimed should not be unduly limited to such specific embodiments.

[0046] Examples of mTOR inhibitors suitable for use in the composition of the present invention include Temsirolimus, Sirolimus, Everolimus, Deforolimus, and Zotarolimus. In some embodiments, the composition of the present invention also includes Tacrolimus.

[0047] Examples of phosphatidylcholine suitable for use in the composition of the present invention include soy phosphatidylcholine (SPC), hydrogenated soy phosphatidylcholine (HSPC), dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC). Examples of phosphatidylglycerol include dimyristoylphosphatidylglycerol (DMPG), distearoylphosphatidylglycerol (DSPG), dipalmitoylphosphatidylglycerol (DMPG).

[0048] Examples of guggulsterol derivatives suitable for use in the composition of the present invention include guggulsteryl laurate, guggulsteryl myristate, guggulsteryl palmitate, guggulsteryl stearate, guggulsteryl oleate, guggulsteryl linoleate, guggulsteryl linoleneate.

[0049] In some embodiments, the composition of the present invention comprises antioxidants and or stabilizers. Examples of antioxidants suitable for use in the composition of present invention include, alpha-tocopherol (Vitamin E), alpha-tocopherol polyethylene glycol succinate (TPGS), ascorbic acid, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), sodium metabisulfite (SMB), propyl gallate, cysteine, citric acid.

[0050] In some embodiments, the composition of the present invention contains buffers. Examples of base or buffer include but are not limited to sodium citrate, sodium succinate, sodium phosphate, sodium acetate, sodium hydroxide, saline, and the like.

[0051] The present invention provides compositions comprising mTOR inhibitors and delivering such compositions to a subject, e.g., a human subject. Any suitable amount of mTOR inhibitors sufficient to produce a desired effect, e.g., a therapeutic effect, can be used. In preferred embodiments, suitable amounts of mTOR inhibitors are those amounts that can be suitably incorporated into a suspension, solution, nanoparticles, of the present invention.

[0052] In some embodiments, a composition according to the present invention comprises homogenous suspension, liposomes, micelles, vesicles, nanoparticles that have a mean diameter of about 5 microns or less, while in some embodiments, homogenous suspension, liposomes, micelles, vesicles, nanoparticles that have a diameter of about 1 micron or less. In some embodiments, the homogenous suspension, liposomes, micelles, vesicles, nanoparticles that have a diameter of about 500 nm or less, while in some embodiments, the homogenous suspension, liposomes, micelles, vesicles, nanoparticles that have a diameter of about 200 nm or less. In some preferred embodiments, homogenous suspension, liposomes, micelles, vesicles, nanoparticles that have a diameter of about 100 nm or less.

[0053] The present invention is not limited to any form of composition for example, in some embodiments, a composition according to the present invention is in a lyophilized form. In some embodiments, the composition further comprises a cryoprotectant. In certain preferred embodiments, the cryoprotectant comprises one or more sugars, while in particularly preferred embodiments; the one or more sugars comprise sucrose, lactose, glucose, dextrose, trehalose, maltose, mannitol, and / or sorbitol. In some embodiments of present invention, the cryoprotectant comprises glycine and polyvinylpyrrolidone (PVP). The percentage of sugar in the composition may range from about 10% to about 90%.

[0054] In some embodiments, the lyophilized form is reconstituted with a suitable vehicle to achieve desired mTOR inhibitor concentration. The vehicle for reconstitution includes but not limited to water for injection, sodium chloride solution, dextrose solution, or any pharmaceutically acceptable buffer. The desired mTOR inhibitor concentration upon reconstitution is in between 0.5 mg / mL to about 20 mg / mL. preferably about 1 mg / mL to about 10 mg / mL or preferable to about 1 mg / mL to about 5 mg / mL.

[0055] In some embodiments, the reconstituted product composition is further diluted to desired mTOR inhibitor concentration for administration. The desired mTOR inhibitor concentration is either fixed concentration or concentration for administration based on the body weight of the subject. The vehicle for dilution includes but not limited to 0.9% saline and 5% dextrose solution or any pharmaceutically acceptable vehicle for administration. In some embodiments, the pH of the composition of the invention ranges from about 2 to about 11 , preferably having pH of about 3 to about 8, and more preferably having pH of about 3.5 to pH 8.0.

[0056] In some embodiments, the composition of the present invention contains about 4% to about 100% by weight of total lipid, preferably about 5% to about 60% by weight of total lipid or more preferably about 8% to about 50% by weight of total lipid.

[0057] In some embodiments, the composition of the present invention contains mTOR inhibitor and total lipid(s) in weight-to- weight ratio between 1 :1 and 1:80; for example, in between 1:1 and 1: 10 weight ratio or in between 1: 1 and 1 :20 weight ratio or in between 1 :1 and 1 :30 weight ratio, or in between 1: 1 and 1 :40 weight ratio or in between 1 : 1 and 1:50 weight ratio, or in between 1 : 1 and 1 :60 weight ratio, or in between 1 : 1 and 1 :70 weight ratio or in between 1 : 1 and 1 :80 weight ratio. In preferred embodiments, the composition of present invention contains mTOR inhibitors and total lipid(s) in weight-to- weight ratio between 1 :5 and 1 :50. As used herein, the term “in between” is inclusive of the limits of the range. For example, a weight-to-weight ratio in between 1 :5 and 1:50 weight ratio includes weight ratio of 1 :5 and 1:50.

[0058] In some embodiments, the composition of the present invention contains mTOR inhibitor and phosphatidylcholine in weight-to-weight ratio between 1 : 1 and 1 :80; for example, in between 1 : 1 and 1: 10 weight ratio or in between 1 : 1 and 1 :20 weight ratio or in between 1 : 1 and 1 :30 weight ratio, or in between 1: 1 and 1 :40 weight ratio or in between 1 : 1 and 1:50 weight ratio, or in between 1 : 1 and 1 :60 weight ratio, or in between 1 : 1 and 1 :70 weight ratio or in between 1 : 1 and 1 :80 weight ratio. In preferred embodiments, the composition of present invention contains mTOR inhibitors and total lipid(s) in weight-to-weight ratio between 1 :5 and 1 :50. As used herein, the term “in between” is inclusive of the limits of the range. For example, a weight-to-weight ratio in between 1 :5 and 1:50 weight ratio includes weight ratio of 1 :5 and 1:50. Examples of phosphatidylcholine include soy phosphatidylcholine (SPC), hydrogenated soy phosphatidylcholine (HSPC), dimyristoylphosphatidylcholine (DMPC), dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC).

[0059] In some embodiments, the composition of the present invention contains mTOR inhibitor and phosphatidylglycerol in weight-to-weight ratio between 1: 1 and 1:80; for example, in between 1: 1 and 1 :10 weight ratio or in between 1: 1 and 1:20 weight ratio or in between 1 : 1 and 1 :30 weight ratio, or in between 1 : 1 and 1 :40 weight ratio or in between 1 : 1 and 1 :50 weight ratio, or in between 1: 1 and 1 :60 weight ratio, or in between 1: 1 and 1:70 weight ratio or in between 1 :1 and 1:80 weight ratio. In preferred embodiments, the composition of present invention contains mTOR inhibitors and total lipid(s) in weight-to- weight ratio between 1:5 and 1:50. As used herein, the term “in between” is inclusive of the limits of the range. For example, a weight-to-weight ratio in between 1 : 5 and 1 : 50 weight ratio includes weight ratio of 1 :5 and 1 :50. Examples of phosphatidylglycerol include dimyristoylphosphatidylglycerol (DMPG), distearoylphosphatidylglycerol (DSPG), dipalmitoylphosphatidylglycerol (DMPG).

[0060] In some embodiments, the composition of the present invention contains mTOR inhibitor and guggulsterol or guggulsterol derivative in weight-to-weight ratio between 1:0.1 and 1: 10; for example, in between 1 :0.1 and 1:0.2 weight ratio or in between 1:0.1 and 1:0.3 weight ratio or in between 1:0.1 and 1:0.4 weight ratio, or in between 1:0.1 and 1:0.5 weight ratio or in between 1: 0.1 and 1:0.6 weight ratio, or in between 1:0.1 and 1:0.7 weight ratio, or in between 1 :0.1 and 1 :0.8 weight ratio or in between 1 :0.1 and 1:0.9 weight ratio, or in between 0.1 and 0.10 weight ratio. As used herein, the term “in between” is inclusive of the limits of the range. For example, a weight-to-weight ratio in between 1:0.1 and 1 :10 weight ratio includes weight ratio of 1:0.1 and 1: 10. Examples of guggulsterol derivatives include guggulsteryl laurate, guggulsteryl myristate, guggulsteryl palmitate, guggulsteryl stearate, guggulsteryl oleate, guggulsteryl linoleate, guggulsteryl linoleneate.

[0061] In some embodiments, the composition of the present invention contains mTOR inhibitor and mixture of phosphatidylcholine and guggulsterol or guggulsterol derivative. The weight-to-weight ratio of mTOR inhibitors and the mixture of phosphatidylcholine and guggulsterol or guggulsterol derivative is in between 1:1 and 1 :80; for example, in between 1:1 and 1: 10 weight ratio or in between 1: 1 and 1:20 weight ratio or in between 1: 1 and 1:30 weight ratio, or in between 1 : 1 and 1 :40 weight ratio or in between 1 : 1 and 1:50 weight ratio, or in between 1 : 1 and 1 :60 weight ratio, or in between 1 : 1 and 1 :70 weight ratio or in between 1:1 and 1 :80 weight ratio. In preferred embodiments, the composition of present invention contains mTOR inhibitors and mixture of phosphatidylcholine and guggulsterol or guggulsterol derivative in weight-to- weight ratio between 1 :5 and 1:50. As used herein, the term “in between” is inclusive of the limits of the range. For example, a weight-to- weight ratio in between 1 : 5 and 1 :50 weight ratio includes weight ratio of 1 : 5 and 1:50.

[0062] In some embodiments, the composition of the present invention contains mTOR inhibitor and mixture of phosphatidylglycerol and guggulsterol or guggulsterol derivative. The weight-to-weight ratio of mTOR inhibitors and the mixture of phosphatidylglycerol and guggulsterol or guggulsterol derivative is in between 1: 1 1 :80; for example, in between 1 : 1 and 1:10 weight ratio or in between 1: 1 and 1:20 weight ratio or in between 1:1 and 1 :30 weight ratio, or in between 1 :1 and 1:40 weight ratio or in between 1 : 1 and 1 :50 weight ratio, or in between 1 : 1 and 1 :60 weight ratio, or in between 1 : 1 and 1 :70 weight ratio or in between 1 : 1 and 1 :80 weight ratio. In preferred embodiments, the composition of present invention contains mTOR inhibitors and mixture of phosphatidylglycerol and guggulsterol or guggulsterol derivative in weight-to-weight ratio between 1:5 and 1:50. As used herein, the term “in between” is inclusive of the limits of the range. For example, a weight-to-weight ratio in between 1 : 5 and 1 : 50 weight ratio includes weight ratio of 1 : 5 and 1:50.

[0063] In some embodiments, the composition of the present invention contains mTOR inhibitor and mixture of phosphatidylcholine, phosphatidylglycerol and guggulsterol or guggulsterol derivative. The weight-to-weight ratio of mTOR inhibitors and the mixture of phosphatidylcholine, phosphatidylglycerol, and guggulsterol or guggulsterol derivative is in between 1 : 1 and 1:80; for example, in between 1 : 1 and 1: 10 weight ratio or in between 1 : 1 and 1:20 weight ratio or in between 1:1 and 1:30 weight ratio, or in between 1: 1 and 1 :40 weight ratio or in between 1: 1 and 1:50 weight ratio, or in between 1: 1 and 1 :60 weight ratio, or in between 1 :1 and 1:70 weight ratio or in between 1: 1 and 1 :80 weight ratio. In preferred embodiments, the composition of present invention contains mTOR inhibitors and mixture of phosphatidylcholine, phosphatidylglycerol and guggulsterol or guggulsterol derivative in weight-to-weight ratio between 1 :5 and 1:50. As used herein, the term “in between” is inclusive of the limits of the range. For example, a weight-to-weight ratio in between 1 : 5 and 1 : 50 weight ratio includes weight ratio of 1 :5 and 1:50.

[0064] In some embodiments, the compositions of the present invention include antioxidants. Examples of antioxidants include but are not limited to alpha-tocopherol (Vitamin E), alpha- tocopherol polyethylene glycol succinate (TPGS), ascorbic acid, butylated hydroxytoluene (BHT), butylated hydroxyanisole (BHA), sodium metabisulfite (SMB), propyl gallate, cysteine, citric acid, etc.

[0065] In some embodiments, the compositions of the present invention contain mTOR inhibitor about 0.1% to about 90% of the total weight, preferably about 0.5% to about 75% of the total weight, more preferably about 1% to about 50% of the total weight.

[0066] In some embodiments, this invention provides compositions comprising mTOR inhibitors and lipid(s) and administering said mTOR inhibitors and lipid(s) to a human subject. In some embodiments, the administering of said composition is through a parenteral route of administration. In some embodiments, the administering of said composition is through a rectal route of administration.

[0067] In some embodiments, the administration of said composition comprising mTOR inhibitors and lipid(s) is in the form of suspension, solution, nanoparticles, vesicles, emulsion.

[0068] In some embodiments, the composition of present invention after parenteral administration provides a similar pharmacokinetic profile compared to the reference product when equal dose of temsirolimus is administered. The said pharmacokinetic profile includes concentrations of temsirolimus and its metabolite, sirolimus in blood at different time points after said administration of the composition of present invention and the reference product. Example of reference product include commercially available solvent based temsirolimus injection product comprising dehydrated alcohol, propylene glycol, polysorbate 80, polyethylene glycol 400 (PEG400).

[0069] In a more preferred embodiment, the composition of the present invention in lyophilized powder form is reconstituted with water for injection and diluted with 0.9% sodium chloride solution to a desired concentration. Similarly, the commercially available reference product, temsirolimus injection comprising dehydrated alcohol (39.5% w / v) and propylene glycol (50.3%) is diluted with the diluent provided with the said reference product comprising polysorbate 80 (40.0% w / v), PEG400 (42.8% w / v), and 19.9% w / v) and further diluted with 0.9% sodium chloride solution to a desired concentration. In the said preferred embodiment, equal doses of temsirolimus from said diluted composition of present invention and the said diluted reference product provides similar pharmacokinetic profile after intravenous administration. The said pharmacokinetic profile includes concentrations of temsirolimus and its metabolite, sirolimus in blood at different time point intervals after said intravenous administration of the diluted composition of present invention and the reference product.

[0070] In some embodiments, the composition of present invention after spiking in whole human blood provides a similar temsirolimus concentration compared to the reference product when equal dose of temsirolimus is spiked and incubated at 37 °C. Example of reference product include commercially available solvent based temsirolimus injection product comprising dehydrated alcohol, propylene glycol, polysorbate 80, polyethylene glycol 400 (PEG400).

[0071] In a more preferred embodiment, the composition of the present invention in lyophilized powder form is reconstituted with water for injection and diluted with 0.9% sodium chloride solution to a desired concentration. Similarly, the commercially available reference product, temsirolimus injection comprising dehydrated alcohol (39.5% w / v) and propylene glycol (50.3%) is diluted with the diluent provided with the said reference product comprising polysorbate 80 (40.0% w / v), PEG400 (42.8% w / v), and 19.9% w / v) and further diluted with 0.9% sodium chloride solution to a desired concentration. In the said preferred embodiment, equal doses of temsirolimus from said diluted composition of present invention and the said diluted reference product provides comparable temsirolimus concentration after spiking in whole human blood and incubating at 37 °C.

[0072] In some embodiments, the composition of present invention after spiking into human plasma provides a similar temsirolimus concentration compared to the reference product when equal dose of temsirolimus is spiked and incubated at 37 °C. Example of reference product include commercially available solvent based temsirolimus injection product comprising dehydrated alcohol, propylene glycol, polysorbate 80, polyethylene glycol 400 (PEG400).

[0073] In a more preferred embodiment, the composition of the present invention in lyophilized powder form is reconstituted with water for injection and diluted with 0.9% sodium chloride solution to a desired concentration. Similarly, the commercially available reference product, temsirolimus injection comprising dehydrated alcohol (39.5% w / v) and propylene glycol (50.3%) is diluted with the diluent provided with the said reference product comprising polysorbate 80 (40.0% w / v), PEG400 (42.8% w / v), and 19.9% w / v) and further diluted with 0.9% sodium chloride solution to a desired concentration. In the said preferred embodiment, equal doses of temsirolimus from said diluted composition of present invention and the said diluted reference product provides comparable temsirolimus concentration after spiking in human plasma and incubating at 37 °C.

[0074] The composition of the present invention may be administered in any dosage form and via any system that delivers the active compound mTOR inhibitors in vivo. In some embodiments, a composition of the present invention is delivered in a dosage form selected from parenteral and rectal form. In some embodiments, the composition is formulated into a desired dosage form to achieve immediate release profile, extended-release profile, or delayed release profile in vivo upon administration.

[0075] EXPERIMENTAL EXAMPLES

[0076] EXAMPLE 1

[0077] Soy phosphatidylcholine (1.2 g) was taken in disodium succinate solution in water (2.16 mg / L, pH 6.0; 25 mL) and subjected to high pressure hand homogenization four times. Temsirolimus (30 mg) was added the high-pressure homogenization was continued until desired particle size is achieved. Sucrose (2.25 g) was dissolved in disodium succinate solution (5 mL) and added to temsirolimus, stirred, and filtered through 0.2 p filter. The particle size was determined using Nicomp particle sizer 380. The mean volume weighting diameter amounted to less than 200 nm. Particle size distribution: Mean: 46.2 nm, D90: 95.9 nm, D99: 196.8 nm.

[0078] EXAMPLE 2

[0079] Soy phosphatidylcholine (1.16 g) and guggulsteryl laurate (40.2 mg) were taken in 0.2% sodium citrate solution (25 mL) in water and subjected to high pressure hand homogenization four times. Temsirolimus (60 mg) was added the high-pressure homogenization was continued until desired particle size is achieved. Sucrose (2.25 g) was dissolved in 0.2% sodium citrate solution (5 mL) and added to Temsirolimus-SPC-guggul laurate suspension, stirred, and filtered through 0.2 p filter and lyophilized. The particle size was determined using Nicomp particle sizer 380. The mean volume weighting diameter amounted to less than 200 nm. Particle size distribution: Mean: 34.6 nm, D90: 67.5 nm, D99: 131.4 nm.

[0080] EXAMPLE 3

[0081] Soy phosphatidylcholine (2.0 g) was taken in 0.2% sodium citrate solution (25 mL) in water (40 mL) and subjected to high pressure hand homogenization four times. Temsirolimus (100 mg) was added the high-pressure homogenization continued until desired particle size is achieved. Sucrose (3.75 g) was dissolved in (10 mL) and added to SPC-Temsirolimus suspension, stirred, and filtered through 0.2 p filter and lyophilized. The particle size was determined using Nicomp particle sizer 380. The mean volume weighting diameter amounted to less than 200 nm. Particle size distribution: Mean: 18.5 nm, D90: 42.5 nm, D99: 89.9 nm.

[0082] EXAMPLE 4

[0083] Soy phosphatidylcholine (4.0 g) was taken in 0.2% sodium citrate solution (80 mL) and subjected to high pressure hand homogenization four times. Temsirolimus (200 mg) was added the high-pressure homogenization was continued until desired particle size is achieved. Sucrose (7.5 g) was dissolved in 0.2% sodium citrate solution (7.5%, 20 mL) was added to SPC-Temsirolimus suspension, stirred, and filtered through 0.2 p filter and lyophilized. The particle size was determined using Nicomp particle sizer 380. The mean volume weighting diameter amounted to less than 200 nm. Particle size distribution: Mean: 32.7 nm, D90: 90.8 nm, D99: 176.9 nm.

[0084] EXAMPLE 5

[0085] Soy phosphatidylcholine (1.0 g) was taken in 0.2% sodium citrate solution (20 mL) and subjected to high pressure hand homogenization four times. Sirolimus (25 mg) was added the high-pressure homogenization was continued until desired particle size is achieved. Sucrose (1.875 g) was dissolved in 0.2% sodium citrate solution (7.5%, 5 mL) was added to SPC-Temsirolimus suspension, stirred, and filtered through 0.2 p filter and lyophilized. The particle size was determined using Nicomp particle sizer 380. The mean volume weighting diameter amounted to less than 200 nm. Particle size distribution: Mean: 38.5 nm, D90: 76.3 nm, D99: 156.8 nm.

[0086] EXAMPLE 6

[0087] Soy phosphatidylcholine (4.0 g) was taken in 0.2% sodium citrate solution (80 mL) and subjected to high pressure hand homogenization four times. Temsirolimus (100 mg) was added the high-pressure homogenization continued until desired particle size is achieved. Sucrose (7.5 g) was dissolved in 0.2% sodium citrate solution (7.5%, 20 mL) was added to SPC- Temsirolimus suspension, stirred, and filtered through 0.2 p filter and lyophilized. The particle size was determined using Nicomp particle sizer 380. The mean volume weighting diameter amounted to less than 200 nm. Particle size distribution: Mean: 29.0 nm, D90: 64.0 nm, D99: 128.1 nm.

[0088] EXAMPLE 7

[0089] Soy phosphatidylcholine (5.0 g), Temsirolimus (250 mg), Monosodium citrate (200 mg), and Sucrose (24.05 g) were mixed in water for injection under stirring using overhead stirrer. The resulting suspension was homogenized using high-pressure homogenization until desired particle size is achieved. The volume of the suspension was adjusted to desired volume and filtered through 0.2 p filter and lyophilized. The particle size was determined using Nicomp particle sizer 380. The mean volume weighing diameter amounted to less than 200 nm.

[0090] EXAMPLE 8

[0091] Acute Intravenous Toxicity Study of Temsirolimus Lipid Suspension for Injection in Wistar Rats

[0092] The temsirolimus lipid suspension (prepared according to Example 6) was reconstituted with 0.9% saline and diluted at dose levels of 0, 5, 10, and 25 mg / kg temsirolimus / body weight was administered intravenously to Wistar rats (10 animals / dose, 5 males and 5 females). The rats in the control group (0 mg dose level) were similarly treated with n-saline intravenously at the equivalent volume as per kg body weight. The animals were monitored for clinical signs, body weight, mortality, necropsy (gross pathology) and histopathology. The results suggested that temsirolimus lipid suspension has no serious adverse effects at all dose levels tested. The Maximum Tolerated Dose (MTD) was found at 25 mg / kg (high dose) when temsirolimus Lipid Suspension for Injection was administered to Wistar rats by the intravenous route Dose Levels: 0, 5, 10, 25 mg / kg.

[0093] EXAMPLE 9

[0094] Acute Intravenous Toxicity Study of Temsirolimus Lipid Suspension for Injection in Swiss Albino Mice

[0095] The temsirolimus lipid suspension (prepared according to Example 6) was reconstituted with 0.9% saline and diluted at dose levels of 0, 10, 20, and 50 mg / kg temsirolimus / body weight was administered intravenously to Swiss Albino mice (10 animals / dose, 5 males and 5 females). The mice in the control group (0 mg dose level) were similarly treated with n-saline intravenously at the equivalent volume as per kg body weight. The animals were monitored for clinical signs, body weight, mortality, necropsy (gross pathology) and histopathology. The results suggested that temsirolimus lipid suspension has no serious adverse effects at all dose levels tested. The Maximum Tolerated Dose (MTD) was found at 50 mg / kg (high dose) when temsirolimus Lipid Suspension for Injection was administered to Swiss Albino mice by the intravenous route Dose Levels: 0, 10, 20, and 50 mg / kg.

[0096] EXAMPLE 10

[0097] Stability of Infusion Preparations (0.1 mg / mL) at room temperature

[0098] The temsirolimus lipid suspension (prepared according to Example 6) was reconstituted and diluted with 0.9% NaCl to recommended concentration for infusion at 0.1 mg temsirolimus / mL. The infusion preparations were incubated at room temperature for up to 4 hrs. The concentrations of temsirolimus were monitored with a validated LC-MS / MS method at 0.5 and 4 hrs., respectively. There is little change in concentration of temsirolimus over a period of 4 hrs at room temperature.

[0099] Table 1

[0100] EXAMPLE 11

[0101] Comparable Partitioning of Temsirolimus in red blood cells (RBC) and plasma following incubation of Temsirolimus Lipid Suspension for Injection (Test Product) or Temsirolimus Injection (Reference Product) in human whole blood at 37 °C.

[0102] Temsirolimus Lipid Suspension for Injection (Test Product, T): The lyophilized product (prepared according to Example 6) was reconstituted with water for injection to achieve 1 mg / mL temsirolimus concentration and further diluted with 0.9% sodium chloride to achieve 50 pg / mL temsirolimus concentration.

[0103] Temsirolimus Injection (Reference Product, R): The reference product vial comprising dehydrated alcohol (39.5% w / v) and propylene glycol (50.3%) was diluted with the diluent provided with the said reference product comprising polysorbate 80 (40.0% w / v), PEG400 (42.8% w / v), and 19.9% w / v) and further diluted with 0.9% sodium chloride solution to achieve 50 pg / mL temsirolimus concentration.

[0104] Whole blood was spiked with either Test or Reference (50 pg / mL) to achieve Temsirolimus concentration of approx. 1 pg / mL. This level is close to the maximum concentration (Cmax) following a dose of 25 mg infused over 0.5 -1 hrs. period in patients. Following the incubation at 37 °C, RBC were separated, and the levels of temsirolimus in RBC and plasma were measured with a validated LC-MS / MS method, respectively. The results showed that there were no statistical differences in RBC partitioning of temsirolimus between Test and Reference. Table 2

[0105] EXAMPLE 12

[0106] Comparable levels of free (plasma protein unbound) Temsirolimus for Temsirolimus Lipid Suspension for Injection (Test Product) or Temsirolimus Injection (Reference Product) at 37 °C.

[0107] Temsirolimus Lipid Suspension for Injection (Test Product, T): The lyophilized product (prepared according to Example 6) was reconstituted with water for injection to achieve 1 mg / mL temsirolimus concentration and further diluted with 0.9% sodium chloride to achieve 50 pg / mL temsirolimus concentration.

[0108] Temsirolimus Injection (Reference Product, R): The reference product vial comprising dehydrated alcohol (39.5% w / v) and propylene glycol (50.3%) was diluted with the diluent provided with the said reference product comprising polysorbate 80 (40.0% w / v), PEG400 (42.8% w / v), and 19.9% w / v) and further diluted with 0.9% sodium chloride solution to achieve 50 pg / mL temsirolimus concentration.

[0109] Above diluted Test or Reference formulations were spiked into human plasma to achieve approx. 1 pg / mL final concentration. The plasma samples were incubated for up to 4 hrs. at 37 °C and level of free (protein unbound) temsirolimus in plasma was measured at 0.5 and 4 hrs. time points with a validated LC-MS / MS method. The results showed that the plasma level of free temsirolimus from Test and Reference formulations are comparable. Table 3

[0110] EXAMPLE 13

[0111] Comparative Pharmacokinetics of Temsirolimus Lipid Suspension for Injection (Test Product) and Temsirolimus Injection (Reference Product) in ICR (CD-I) Mice

[0112] Temsirolimus Lipid Suspension for Injection (Test Product, T): The lyophilized product (prepared according to Example 6) was reconstituted with water for injection to achieve 1 mg / mL temsirolimus concentration and used without further dilution.

[0113] Temsirolimus Injection (Reference Product, R): The reference product vial comprising dehydrated alcohol (39.5% w / v) and propylene glycol (50.3%) was diluted with the diluent provided with the said reference product comprising polysorbate 80 (40.0% w / v), PEG400 (42.8% w / v), and 19.9% w / v) and further diluted with 0.9% sodium chloride solution to achieve 1 mg / mL temsirolimus concentration.

[0114] A single equal dose of Test Product (T) or Reference Product (R) was administered intravenously in ICR (CD-I) mice.

[0115] Result: The pharmacokinetics of temsirolimus and its main metabolite, Sirolimus, following a single intravenous dose administration from temsirolimus Lipid Suspension for Injection (Test) or from solvent-based temsirolimus Injection (Reference) at a dose level of 10 mg / kg body weight was evaluated in ICR (CD-I) mice. Following equal dose administration, blood was collected over a 24-hr period with one blood sample from each mouse (3 mice per time point). Blood levels of temsirolimus and sirolimus in each sample were analyzed by an HPLC method. The blood concentrations of temsirolimus and sirolimus were calculated from a standard curve and shown in FIG. 1 and FIG. 2, respectively. The blood concentrations of both temsirolimus and sirolimus from Test and Reference formulations are found comparable. REFERENCES

[0116] 1. Bhaskar, P.T., and Hay, N. Developmental Cell, 2007, 12, 487-502.

[0117] 2. Lench, N.J., Macadam, R., Markham, A.F. Human genetics, 1997, 99, 547-549.

[0118] 3. Palavra, F., Robalo, C., Reis, F. Oxidative Medicine and Cellular Longevity, 2007, Article ID 9820181, 11 pages.

[0119] 4. Viana, S.D., Reis, F., Alves, R., Oxidative Medicine and Cellular Longevity, 2018, Article ID 3693625, 17 pages.

[0120] 5. Kelchtermans, J., Chang, J., Glaberson, W., DeFreitas, M., Alba-Sandoval, M., Chandar, J. J. Pediatr Pharmacol Ther, 2020, 25, 459-464.

[0121] 6. Feng, Y., Chen, X., Cassady, K., Zou, Z., Yang, S., Wang, Z., Zhang, X. Frontiers in Oncology, 2021, 10, Article ID 611690.

[0122] 7. Rodriguez-Rodriguez, A.E., Donate-Correa, J., Rovira, J., Cuesto, G., Luis-Ravelo, D., Fernandes, M.X., Acevedo- Arozena, A., Diekman, F., Acebes, A., Torres, A., Porrini E. Am. J. Transplant, 2019, 19, 3240-3249.

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Claims

CLAIMSWhat is claimed is:

1. A composition comprising mTOR inhibitor and at least one lipid and / or guggulsterol or guggulsterol derivative.

2. The composition of claim 1 , wherein said mTOR inhibitor is selected from a group consisting of sirolimus, everolimus, temsirolimus, deforolimus, zotarolimus, and tacrolimus.

3. The composition of claim 1, wherein said at least one lipid is selected from the group consisting of soy phosphatidylcholine (SPC), hydrogenated soy phosphatidylcholine, dimyristoylphosphatidylcholine (DMPC), dimyristoylphosphatidylglycerol (DMPG), dipalmitoylphosphatidylcholine (DPPC), disteroylphosphatidylglycerol (DSPG), and dipalmitoylphosphatidylglycerol (DMPG).

4. The composition of claim 1 wherein, the guggulsterol derivative is selected from a group consisting of guggulsteryl laurate, guggulsteryl myrstate, guggulsteryl palmitate, guggulsteryl stearate, guggulsteryl oleate, guggulsteryl linoleate, and guggulsteryl linolenate.

5. The composition of claim 1 further contains a cryoprotectant.

6. The composition of claim 5, wherein the cryoprotectant is selected, from lactose, glucose, dextrose, trehalose, maltose, mannitol, and sorbitol.

7. The composition of claim 1 , wherein said composition is in a powder form.

8. The composition of claim 1, wherein said composition is in a suspension form.

9. The composition of claims 1 and 8, the concentration of said mTOR inhibitor in the said suspension is in between 1 mg / mL to 5 mg / mL.

10. The composition of claim 1, wherein the amount of said mTOR inhibitor is about 0.5% to about 50% of the total weight.

11. The composition of claim 1, wherein the weight-to- weight ratio of said mTOR inhibitor and lipid(s) is in between 1 :5 and 1:50.

12. The composition of claim 1, wherein the weight-to- weight ratio of said mTOR inhibitor and guggulsterol or gugguslsterol derivative is in between 1:0.1 and 1: 10.

13. The composition of claims 1, 7, and 8 wherein said composition comprises parenteral administration in a subject.

14. The composition of claims 1 and 13, wherein the said parenteral administration provides comparable pharmacokinetics of mTOR inhibitors upon administration with solvent based temsirolimus injection comprising dehydrated alcohol, propylene glycol, polysorbate 80, polyethylene glycol 400 (PEG400).

15. The composition of claims 13 and 14, wherein said subject is a human.

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