Subcutaneous administration of nanoparticles comprising mtor inhibitor and albumin for treatment of diseases

Abstract

To provide nanoparticle formulations comprising an mTOR inhibitor that do not cause unacceptable toxicological effects upon administration, such as subcutaneous administration.SOLUTION: The present invention provides compositions and devices for subcutaneous administration of compositions comprising nanoparticles comprising an mTOR inhibitor and an albumin. The present application also provides methods of treating diseases by subcutaneously administering to an individual a composition comprising nanoparticles comprising an mTOR inhibitor and an albumin.SELECTED DRAWING: None

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Application No. 62 / 820,842, filed March 19, 2019, entitled "Subcutaneous Administration of Nanoparticles Comprising an mTOR Inhibitor and Albumin for the Treatment of Disease"; and U.S. Provisional Application No. 62 / 820,838, filed March 19, 2019, entitled "Methods and Compositions for the Treatment of Pulmonary Hypertension"; each of which is incorporated herein by reference for all purposes.

[0002] This application relates to compositions, devices, and methods for subcutaneously administering nanoparticles comprising an mTOR inhibitor and albumin. This application further relates to methods of treating an individual, comprising subcutaneously administering a composition comprising nanoparticles comprising an mTOR inhibitor and albumin. [Background technology]

[0003] The mammalian target of rapamycin (mTOR) is a protein kinase known to regulate various cellular processes, including cell survival, proliferation, stress, and metabolism. Many inhibitors of mTOR, such as rapamycin, are effective in treating various disorders, including certain cancers. Many mTOR inhibitors, such as rapamycin, are known to have poor water solubility and therefore require excipients, such as surfactants and solvents. These excipients can cause irritation, inflammation, and reduced efficacy, especially when administered parenterally, such as subcutaneously.

[0004] Therefore, there is a need in the art for an improved nanoparticle formulation that is stable and / or does not cause unacceptable toxic effects when administered, such as by subcutaneous administration.There is also a need to develop a nanoparticle formulation that is comprised of an mTOR inhibitor, that can be dried, such as by lyophilization, and more easily constituted and / or delivered.Finally, there is a need in the art for reducing the risk of mishandling dry compositions before administration.

[0005] The disclosures of all references, patents, patent applications, and published patent applications cited herein are hereby incorporated by reference in their entirety. Summary of the Invention

[0006] The present application provides a method for treating a disease in an individual, comprising subcutaneously administering to the individual a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor and albumin, wherein the amount of the mTOR inhibitor in the pharmaceutical composition is about 0.1 mg / m for each administration. 2 to approximately 10 mg / m 2 In some embodiments, the amount of mTOR inhibitor in the pharmaceutical composition is about 1 mg / m for each administration. 2 to approximately 10 mg / m 2 In some embodiments, the amount of mTOR inhibitor in the pharmaceutical composition is about 5 mg / m 2 The dosage is:

[0007] In some embodiments according to any one of the methods described herein, the pharmaceutical composition further comprises a sugar.

[0008] In some embodiments of any one of the methods described herein, the pharmaceutical composition is administered no more than once a week.In some embodiments, the pharmaceutical composition is administered once a week.In some embodiments, the pharmaceutical composition is administered twice every three weeks.

[0009] In some embodiments of any one of the methods described herein, the disease is cancer. In some embodiments of any one of the methods described herein, the disease is a mitochondrial disease.

[0010] In some embodiments according to any one of the methods described herein, the individual is a human.

[0011] The present application also provides a method for delivering an effective amount of an mTOR inhibitor to a target tissue of an individual, comprising subcutaneously administering a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor and albumin. In some embodiments, the mTOR inhibitor is rapamycin. In some embodiments, the pharmaceutical composition further comprises a sugar. In some embodiments, the pharmaceutical composition comprises a dose of about 0.1 mg / m 2 to approximately 10 mg / m 2 In some embodiments, the target tissue is brain tissue of the individual.

[0012] In some embodiments of any one of the methods described herein, the average diameter of nanoparticles in pharmaceutical composition is about 120nm or less.In some embodiments of any one of the methods described herein, the nanoparticles comprise the mTOR inhibitor coated with albumin.In some embodiments of any one of the methods described herein, the albumin is human albumin.In some embodiments of any one of the methods described herein, the mTOR inhibitor is a limus drug.In some embodiments of any one of the methods described herein, the mTOR inhibitor is rapamycin.

[0013] The present application also provides a pharmaceutical composition suitable for subcutaneous administration to an individual, comprising: a) nanoparticles comprising an mTOR inhibitor and albumin; and b) a sugar. In some embodiments, the sugar is selected from the group consisting of alginate, starch, lactose, pullulan, hyaluronic acid, chitosan, glucose, galactose, mannose, N-acetylglucosamine, sucrose, N-acetyl-D-galactosamine, maltose, or trehalose. In some embodiments, the sugar is sucrose. In some embodiments, the sugar is trehalose. In some embodiments, the concentration of the mTOR inhibitor in the pharmaceutical composition is at least about 5 mg / ml. In some embodiments, the concentration of the mTOR inhibitor in the pharmaceutical composition is at least about 50 mg / ml. In some embodiments, the average diameter of the nanoparticles in the pharmaceutical composition is about 120 nm or less. In some embodiments, the nanoparticles in the pharmaceutical composition comprise an mTOR inhibitor coated with albumin. In some embodiments, the albumin in the pharmaceutical composition is human albumin. In some embodiments, the mTOR inhibitor in the pharmaceutical composition is a limus drug. In some embodiments, the mTOR inhibitor is rapamycin.

[0014] The present application also provides a device for subcutaneously administering to an individual a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor and albumin, the device comprising: a) a drug chamber containing the pharmaceutical composition in dry form, and a solution chamber containing a reconstituted solution; and a removable partition separating the drug chamber and the solution chamber, wherein removal of the partition causes the dry pharmaceutical composition and the reconstituted solution to mix, thereby forming a reconstituted pharmaceutical composition. In some embodiments, the device is a syringe, comprising an injection needle attached to the end of the syringe and a pusher that can eject the reconstituted pharmaceutical composition from the syringe. In some embodiments, the pharmaceutical composition of the device further comprises a sugar. In some embodiments of the device, the mTOR inhibitor is a limus drug. In some embodiments of the device, the mTOR inhibitor is rapamycin.

[0015] The present application also provides kits comprising any of the devices described herein for use in treating a disease. In some embodiments, the kit further comprises instructions for using the kit to treat cancer. In some embodiments, the kit further comprises instructions for using the kit to treat a mitochondrial disease. [Brief explanation of the drawings]

[0016] [Figure 1] Figure 1 shows rapamycin concentrations in whole blood samples taken from rats after subcutaneous (SC) or intravenous (IV) administration of a single dose of nab-rapamycin (ABI-009) between 0 and 24 hours post-administration.

[0017] [Figure 2] Figure 2 shows rapamycin concentrations in whole blood samples taken from rats after subcutaneous (SC) or intravenous (IV) administration of a single dose of nab-rapamycin (ABI-009) between 0 and 168 hours post-dose.

[0018] [Figure 3]Figure 3 shows rapamycin concentrations in whole blood samples taken from rats after subcutaneous (SC) or intravenous (IV) administration of a single dose of nab-rapamycin (ABI-009) between 0 and 24 hours post-administration.

[0019] [Figure 4] FIG. 4 shows the bioavailability of nab-rapamycin (ABI-009) after single dose subcutaneous (subQ) or intravenous (IV) administration in rats, as indicated by the calculated area under the curve (AUC).

[0020] [Figure 5] FIG. 5 shows the concentration of rapamycin in the bone marrow (top) or brain (bottom) of rats 24 or 168 hours after subcutaneous (subQ) or intravenous (IV) administration of a single dose of nab-rapamycin (ABI-009).

[0021] [Figure 6] FIG. 6 shows the concentration of rapamycin in the heart (top) or liver (bottom) of rats 24 or 168 hours after subcutaneous (subQ) or intravenous (IV) administration of a single dose of nab-rapamycin (ABI-009).

[0022] [Figure 7] FIG. 7 shows the concentration of rapamycin in the lungs (top) or pancreas (bottom) of rats 24 or 168 hours after subcutaneous (subQ) or intravenous (IV) administration of a single dose of nab-rapamycin (ABI-009).

[0023] [Figure 8] Figure 8 shows a comparison of the time course of rapamycin concentrations in the brain or whole blood of rats at 24, 72, and 120 hours after administration of a single subcutaneous dose of nab-rapamycin (ABI-009) at doses of 1.7 mg / kg, 9.5 mg / kg, or 17 mg / kg.

[0024] [Figure 9]FIG. 9 shows a comparison of the histopathology scores assessed in the skin of rats between the different treatment groups.

[0025] [Figure 10] Figure 10 shows a representative histogram image of skin from a rat in Group 1 (0.9% saline). Histological lesions are limited to aggregates of mixed inflammatory cells (black arrows) within the subcutaneous tissue (SC). The dermis (D) and epidermis (E) are shown.

[0026] [Figure 11] Figure 11 shows a representative histogram image of skin from a rat in Group 2 (0.9% HSA in saline). Multifocal mixed inflammatory cell aggregates (black arrows) are visible in the subcutaneous tissue (SC). They are unremarkable in the epidermis (E) and dermis (D).

[0027] [Figure 12] Figure 12 shows a representative histogram image of skin from a rat in Group 3 (ABI-009, 1.7 mg / kg). Minimal mixed inflammatory cell infiltrate (black arrow) is visible in the subcutaneous tissue (SC). The epidermis (E) and dermis (D) are shown.

[0028] [Figure 13] Figure 13 shows a representative histogram image of skin from a rat in Group 4 (ABI-009, 5 mg / kg). In the subcutaneous tissue (SC), there is scattered mixed inflammatory cell infiltrate (right arrow) and minimal areas of necrosis (left arrow). The epidermis (E) and dermis (D) are unremarkable.

[0029] [Figure 14] Figure 14 shows a representative histogram image of skin from a rat in Group 4 (ABI-009, 10 mg / kg). Subcutaneous (SC) mixed inflammatory cell infiltrate (right arrow) and areas of necrosis (left arrow) are noted. The epidermis (E) and dermis (D) are unremarkable.

[0030] [Figure 15]FIG. 15 depicts the mean blood concentrations of rapamycin in rats administered ABI-009 at 1.7 mg / kg, 5 mg / kg, or 10 mg / kg.

[0031] [Figure 16] FIG. 16 shows the results of tumor growth in a human hepatocellular carcinoma mouse xenograft model 0 to 15 days after treatment with saline (Group 1), ABI-009 (intravenous route; Group 2), Rapamune (oral administration; Group 3), and ABI-009 (subcutaneous route; Group 4).

[0032] [Figure 17] Figure 17 shows the weight changes of mice in a human hepatocellular carcinoma mouse xenograft model 0 to 15 days after treatment with saline (Group 1), ABI-009 (intravenous route; Group 2), Rapamune (oral administration; Group 3), and ABI-009 (subcutaneous route; Group 4). DETAILED DESCRIPTION OF THE INVENTION

[0033] Detailed Description Provided herein is a method for subcutaneously administering the compositions described herein, such as a composition comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin.In another aspect, provided herein is a method for delivering an effective amount of an mTOR inhibitor (such as rapamycin) to target tissues such as the brain, bone marrow, heart, liver, lung, or pancreatic tissue by subcutaneously administering a composition comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin.In another aspect, provided herein is a method for maintaining the blood concentration of an mTOR inhibitor (such as rapamycin), comprising subcutaneously administering a composition comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin.

[0034] Also provided herein is a method for treating a disease, comprising subcutaneously administering a composition comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin.In some embodiments, the disease is cancer.In some embodiments, individuals with cancer are selected for treatment based on having mTOR activation abnormalities.In some embodiments, the disease is mitochondrial disease.

[0035] Also provided herein are compositions, such as pharmaceutical compositions suitable for subcutaneous administration to individuals, comprising nanoparticles containing an mTOR inhibitor (such as rapamycin) and albumin, as well as methods for administering such compositions.In some aspects, the composition can comprise one or more agents for improving the solubility of the composition in dry form and / or improving the stability of the composition.Additional agents can include sugars.The sugars can be present in an amount effective to improve solubility, such as the dissolution rate after adding the composition in dry form to an aqueous solution, and / or promote the stability of the composition.

[0036] In another aspect, a device for subcutaneously administering the pharmaceutical composition described herein is provided. The device includes a drug chamber containing the pharmaceutical composition in dry form and a solution chamber containing a reconstituted solution. The device further includes a removable partition separating the drug chamber and the solution chamber, wherein removal or activation of the partition causes or allows mixing of the dried pharmaceutical composition and the reconstituted solution, thereby forming a reconstituted pharmaceutical composition. The device may be a syringe and may further include a pusher. After reconstitution of the composition, the device may be capable of or adapted to subcutaneously administer the composition to an individual. Also provided herein is a method for subcutaneously administering a composition containing nanoparticles containing an mTOR inhibitor and albumin using the device described herein.

[0037] definition It is understood that aspects and embodiments of the invention described herein include "consisting of" and / or "consisting essentially of" aspects and embodiments.

[0038] As described herein, albumin can be "associated" with an mTOR inhibitor (such as rapamycin), e.g., a composition includes an albumin-associated mTOR inhibitor. "Associated" or "associated" is used in a general sense herein to refer to albumin affecting the behavior and / or properties of an mTOR inhibitor (such as rapamycin) in an aqueous composition. For example, albumin and an mTOR inhibitor (such as rapamycin) are considered to be "associated" if albumin makes the mTOR inhibitor (such as rapamycin) more easily suspendable in an aqueous solvent compared to a composition that does not contain albumin. As another example, albumin and an mTOR inhibitor (such as rapamycin) are associated if albumin stabilizes the mTOR inhibitor (such as rapamycin) in an aqueous suspension. For example, albumin and an mTOR inhibitor can be present in particles or nanoparticles, as further described herein.

[0039] A general reference to a "composition" can include any pharmaceutical composition described herein.

[0040] As used herein, the term "effective amount" refers to an amount of a compound or composition sufficient to treat a particular disorder, condition, or disease, e.g., to improve, alleviate, relieve, and / or delay one or more of its symptoms. As understood in the art, an "effective amount" may be one or more doses, i.e., single or multiple doses may be required to achieve the desired therapeutic endpoint. An effective amount may be considered in the context of administering one or more therapeutic agents, and a nanoparticle composition (e.g., a composition comprising rapamycin and albumin) may be considered to be an effective amount if, in combination with one or more other agents, a desired or beneficial result can be or is obtained.

[0041] As used herein, "nab" (registered trademark) refers to albumin-bound nanoparticles, and "nab-rapamycin" refers to an albumin-stabilized nanoparticle formulation of rapamycin. Nab-rapamycin is also known as nab-sirolimus, which has been previously described. See, e.g., WO2008 / 109163 A1, WO2014 / 151853, WO2008 / 137148 A2, and WO2012 / 149451 A1, each of which is incorporated herein by reference in its entirety.

[0042] As used herein, "pharmaceutically acceptable" or "pharmacologically compatible" means a substance that is not biologically or otherwise undesirable; e.g., the substance may be incorporated into a pharmaceutical composition administered to a patient without producing significant undesirable biological effects or interacting in a deleterious manner with any other components of the composition in which it is contained. A pharmaceutically acceptable carrier or excipient preferably meets the required standards of toxicology and manufacturing testing and / or is included in the Inactive Ingredients Guide prepared by the U.S. Food and Drug Administration.

[0043] As used herein, "treatment" or "treating" refers to an approach to obtaining a beneficial or desired result, such as a clinical result. For purposes of the present invention, a beneficial or desired clinical result includes, but is not limited to, one or more of the following: alleviating one or more symptoms caused by a disease, alleviating the extent of the disease, stabilizing the disease (e.g., preventing or slowing the progression of the disease), preventing or slowing the spread of the disease (e.g., metastasis), preventing or slowing the recurrence of the disease, reducing the rate of disease recurrence, slowing or slowing the progression of the disease, improving the disease state, providing remission (partial or total) of the disease, reducing the dose of one or more other drugs required to treat the disease, slowing the progression of the disease, improving quality of life, and / or prolonging survival. In some embodiments, treatment reduces the severity of one or more symptoms associated with cancer by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 100%, compared with the corresponding symptoms in the same subject before treatment or compared with the corresponding symptoms in other subjects who have not received treatment.Also, " treatment " includes the alleviation of the pathological effects of cancer.The method of the present invention contemplates any one or more of these aspects of treatment.

[0044] The terms "recurrence," "relapse," and "recurred" refer to the recurrence of cancer or disease after a clinical assessment that the disease had disappeared. A diagnosis of distant metastasis or local recurrence can be considered a recurrence.

[0045] The terms "refractory" or "resistant" refer to a cancer or disease that does not respond to treatment.

[0046] It will be understood that embodiments of the invention described herein include those that "consist of" and / or "consist essentially of" embodiments.

[0047] Reference herein to "about" a value or parameter includes (and describes) a variation about that value or parameter itself. For example, a reference to "about X" includes the description of "X."

[0048] As used herein, a reference to a value or parameter "not" generally means and describes a value or parameter "other than." For example, a reference to a method not being used to treat cancer type X means that the method is used to treat cancer types other than X.

[0049] As used in this specification and the appended claims, the singular forms "a," "or," and "the" include the plural forms unless the context clearly dictates otherwise.

[0050] Subcutaneous administration method Provided herein are methods for subcutaneous administration of a composition, such as a pharmaceutical composition, comprising an mTOR inhibitor, such as rapamycin and albumin.

[0051] In some embodiments, the method provides a method for delivering an effective amount of an mTOR inhibitor (such as rapamycin) to a target tissue, such as the brain, bone marrow, heart, liver, lung, or pancreatic tissue, of an individual, comprising subcutaneously administering a composition, such as a pharmaceutical composition, comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin. In some embodiments, the individual has a tumor in a target tissue, such as the brain.

[0052] In some embodiments, the method provides a method for delivering an effective amount of an mTOR inhibitor (such as rapamycin) to the brain of an individual, comprising subcutaneously administering a composition, such as a pharmaceutical composition, comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin, wherein the dose of the mTOR inhibitor (such as rapamycin) in the nanoparticles to deliver the effective amount of the mTOR inhibitor (such as rapamycin) to the brain is about 0.1 mg / m 2 to approximately 10 mg / m 2 and any quantity of, and values and ranges therein.

[0053] In some embodiments, the method comprises maintaining the blood concentration of an mTOR inhibitor (such as rapamycin) in an individual, the method comprises subcutaneously administering a composition, such as a pharmaceutical composition, comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin. In some embodiments, the blood concentration of the mTOR inhibitor (such as rapamycin) is at least 1 ng / ml, 5 ng / ml, 10 ng / ml, 25 ng / ml, 50 ng / ml, 75 ng / ml, 100 ng / ml, 150 ng / ml, or 200 ng / ml, and any amount and range therein. In some embodiments, the individual has a tumor.

[0054] In some embodiments, the present invention provides a method for treating a disease in an individual, comprising subcutaneously administering to the individual a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor and albumin.In some embodiments, the amount of the mTOR inhibitor in the pharmaceutical composition is about 0.1 mg / m for each administration. 2 to approximately 10 mg / m 2 , for example, about 1 mg / m 2 to approximately 10 mg / m 2 In an exemplary, non-limiting embodiment, the amount of mTOR inhibitor in the pharmaceutical composition is about 5 mg / m for each administration. 2 The dosage is:

[0055] In some embodiments, the amount of mTOR inhibitor in the composition is below that which induces toxicological effects (e.g., effects above clinically acceptable levels of toxicity) when the mTOR inhibitor nanoparticle composition is administered subcutaneously to an individual, or is an amount at which potential side effects are manageable or tolerable. In some embodiments, the toxicological effect is a rash associated with subcutaneous administration of the pharmaceutical composition.

[0056] In some embodiments, the concentration of the mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is between about 0.1 mg / ml and about 100 mg / ml, e.g., about 0.1 mg / ml to about 50 mg / ml, about 0.1 mg / ml to about 50 mg / ml, about 0.1 mg / ml to about 40 mg / ml, about 0.1 mg / ml to about 10 mg / ml, or any amount between about 0.1 mg / ml to about 5 mg / ml, about 5 mg / ml to about 100 mg / ml, about 5 mg / ml to about 50 mg / ml, about 5 mg / ml to about 40 mg / ml, about 7.5 mg / ml to about 100 mg / ml, about 7.5 mg / ml to about 50 mg / ml, about 7.5 mg / ml to about 40 mg / ml, etc., and any value and range therein. In some embodiments, the concentration of the mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is at least any of 5 mg / ml, 7.5 mg / ml, 10 mg / ml, or 20 mg / ml.

[0057] In some embodiments, the effective amount of the mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is in any of the following ranges: about 0.1 mg / m 2 to approximately 5 mg / m 2 , about 5mg / m 2 to approximately 10 mg / m 2 , about 10mg / m 2 to approximately 20 mg / m 2 , about 10 to about 30 mg / m 2 , about 10 to about 45 mg / m 2 , about 10 to about 60 mg / m 2 , about 20 to about 30 mg / m 2 , about 20 to about 45 mg / m 2 , about 20 to about 60 mg / m 2 , about 30 to about 45 mg / m 2 , about 30 to about 60 mg / m 2 , or about 45 to about 60 mg / m 2 In an exemplary, non-limiting embodiment, the effective amount of mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is about 0.1 mg / m 2 to approximately 10 mg / m2 In another exemplary, non-limiting embodiment, the effective amount of mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is between about 1 mg / m 2 to 10 mg / m 2 Between, e.g., 5 mg / m 2 And so on.

[0058] In some embodiments, the dosing frequency for administering the mTOR inhibitor nanoparticle composition (such as a rapamycin / albumin nanoparticle composition) includes, but is not limited to, daily, every 2 days, every 3 days, every 4 days, every 5 days, every 6 days, every week without a break, 3 times in 4 weeks (such as days 1, 8, and 15 of a 28-day cycle), once every 3 weeks, once every 2 weeks, or twice every 3 weeks. In some embodiments, the mTOR inhibitor nanoparticle composition (such as a rapamycin / albumin nanoparticle composition) is administered approximately once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 6 weeks, or once every 8 weeks. In some embodiments, the mTOR inhibitor nanoparticle composition (such as a rapamycin / albumin nanoparticle composition) is administered at least once a week, approximately 1x, 2x, 3x, 4x, 5x, 6x, or 7x (i.e., daily). In some embodiments, the interval between each administration is less than about 6 months, 3 months, 1 month, 20 days, 15 days, 14 days, 13 days, 12 days, 11 days, 10 days, 9 days, 8 days, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day.In some embodiments, the interval between each administration is greater than 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 8 months, or 12 months.In some embodiments, there is no break in the administration schedule.In some embodiments, the interval between each administration is less than about 1 week.

[0059] Administration of the TOR inhibitor nanoparticle composition (such as a rapamycin / albumin nanoparticle composition) can be extended for an extended period, such as from about 1 month up to about 7 years. In some embodiments, the mTOR inhibitor nanoparticle composition (such as a rapamycin / albumin nanoparticle composition) is administered for a period of at least about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, 24, 30, 36, 48, 60, 72, or 84 months.

[0060] The auxiliary substances and adjuvants in any of the compositions described may include, for example, preservatives, wetting agents, suspending agents, flavoring agents, emulsifying agents, and dispersing agents. Prevention of microbial action is generally provided by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, etc. Isotonicity adjusting agents may also be included. Prolonged absorption of injectable dosage forms can be achieved by using agents that delay absorption, such as aluminum monostearate and gelatin. Auxiliaries may also include wetting agents, emulsifying agents, pH buffering agents, and antioxidants, such as citric acid, sorbitan monolaurate, triethanolamine oleate, butylated hydroxytoluene, etc.

[0061] Treatment with any dosing regimen, such as the exemplary dosing regimens described above, can be repeated multiple cycles (1, 2, 3, 4, 5, 6 or more cycles, e.g., about 1 to 10 cycles, 1 to 7 cycles, 1 to 5 cycles, 1 to 4 cycles, 1 to 3 cycles, etc.). In some embodiments, a particular dosing regimen is repeated for at least 2, 3 or more cycles, etc. In some embodiments, treatment with a particular dosing regimen is repeated continuously (i.e., without intervals) for at least 2, 3 or more cycles.

[0062] In some embodiments, there is an interval between two consecutive cycles. In some embodiments, the interval is at least about 1, 2, 3, or 4 weeks. In some embodiments, the interval is at least about 1, 2, 3, 4, 5, 6 months or more. In some embodiments, the interval is approximately the period during which the individual can gain weight (e.g., the individual has a body weight of about or at least about 90%, 92%, 95%, 97% of the body weight before the start of treatment after the interval).

[0063] In some embodiments, the pharmaceutical composition is administered only once.

[0064] Device for subcutaneous administration This application demonstrates that subcutaneous administration of drugs, such as mTOR inhibitor / albumin nanoparticles, exhibits better tolerability and bioavailability characteristics compared to intravenous administration. In some embodiments, compositions comprising nanoparticles comprising an mTOR inhibitor and albumin can be stored in a dry form, such as a lyophilized form. To prepare the dry composition for administration, reconstitution with an aqueous solution, such as water, is required. One aspect of the application provides a device that holds a stable, fixed-dose dry composition in close proximity to a reconstitution solution. The device includes a partition that allows predictable and reproducible reconstitution of the dry composition with limited manipulation, followed by subcutaneous administration of the composition using the device. This device improves handling of the dry composition, for example, by reducing the time required for handling the composition, reducing the opportunity for user error, and ensuring the reproducibility and consistency of the reconstitution process. These advantages are achieved, for example, by eliminating the steps of manually adding the reconstitution solution to the dry composition and completely eliminating the steps of adding the reconstituted composition to a syringe.

[0065] Thus, provided herein is a device for subcutaneously administering compositions, such as pharmaceutical compositions, comprising nanoparticles containing an mTOR inhibitor and albumin. The device described herein is particularly suitable for subcutaneously administering compositions in dry form by sequentially reconstituting the dry form of the composition and then administering the reconstituted composition. In one aspect, the device comprises a drug chamber containing a dry form of a pharmaceutical composition, such as those described herein, such as a lyophilized form of a pharmaceutical composition, and a solution chamber containing a reconstituted solution. In another aspect, the device comprises a partition separating the drug chamber and the solution chamber. Removal or activation of the partition allows and / or causes mixing of the dried pharmaceutical composition and the reconstituted solution, thereby forming a reconstituted pharmaceutical composition. The reconstituted pharmaceutical composition may be suitable for subcutaneous administration to an individual, such as a human. In some embodiments, the device is a syringe comprising a solution chamber and a drug chamber. In some embodiments, the syringe further comprises a pusher that can expel the reconstituted solution from the device. In some embodiments, the syringe further comprises a needle attached to the end of the syringe, such as a hypodermic needle suitable for subcutaneous administration.

[0066] Further provided herein is a composition for subcutaneous administration contained within a device, wherein the composition comprises a lyophilized form of a pharmaceutical composition, and wherein the device comprises a solution chamber containing a reconstitution solution, and a partition separating the drug chamber (containing the pharmaceutical composition) from the reconstitution solution, wherein removal or actuation of the partition allows and / or causes mixing of the dried pharmaceutical composition and the reconstitution solution, thereby forming the reconstituted pharmaceutical composition.

[0067] The divider of the device, in some embodiments, can include a guard to prevent unintentional removal or activation of the divider. In some embodiments, the guard is removed from the device prior to removal or activation of the divider. In some embodiments, activation of the guard allows removal or activation of the divider.

[0068] In some embodiments, the device is a syringe including a needle suitable for injection. In some embodiments, the device is a syringe adapted to mate with a needle suitable for injection. Pressing the pusher of the syringe expels the reconstituted syringe through the needle.

[0069] In some embodiments, a device is provided for subcutaneously administering a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin, the device comprising a drug chamber containing a dry form of the pharmaceutical composition, a solution chamber containing a reconstitution solution, and a removable partition separating the drug chamber and the solution chamber, wherein removal of the partition causes mixing of the dry pharmaceutical composition and the reconstitution solution, thereby forming a reconstituted pharmaceutical composition, and wherein the dose of the mTOR inhibitor (such as rapamycin) in the nanoparticles is any of about 0.2 mg to about 100 mg, about 0.2 mg to about 10 mg, about 10 mg to about 20 mg, about 20 mg to about 30 mg, about 30 mg to about 40 mg, about 40 mg to about 50 mg, about 50 mg to about 60 mg, about 60 mg to about 70 mg, about 70 mg to about 80 mg, about 80 mg to about 90 mg, and about 90 mg to about 100 mg, each inclusive.

[0070] In some embodiments, a device is provided for subcutaneously administering a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin, the pharmaceutical composition optionally comprising a sugar, the device comprising a drug chamber containing the pharmaceutical composition in dry form, a solution chamber containing a reconstitution solution, and a removable partition separating the drug chamber and the solution chamber, wherein removal of the partition results in mixing of the dry pharmaceutical composition with the reconstitution solution, thereby forming a reconstituted pharmaceutical composition, and wherein the dose of the mTOR inhibitor (such as rapamycin) in the nanoparticles is any of about 0.2 mg to about 100 mg, about 0.2 mg to about 10 mg, about 10 mg to about 20 mg, about 20 mg to about 30 mg, about 30 mg to about 40 mg, about 40 mg to about 50 mg, about 50 mg to about 60 mg, about 60 mg to about 70 mg, about 70 mg to about 80 mg, about 80 mg to about 90 mg, and about 90 mg to about 100 mg, each inclusive. In some embodiments, the sugar is selected from the group consisting of alginate, starch, lactose, pullulan, hyaluronic acid, chitosan, glucose, galactose, mannose, N-acetylglucosamine, sucrose, N-acetyl-D-galactosamine, maltose, or trehalose.

[0071] Also provided herein is a method for subcutaneously administering the compositions described herein using the devices described herein. In a non-limiting exemplary embodiment, the composition comprises a dry form of a pharmaceutical composition comprising nanoparticles comprising rapamycin and albumin. In a non-limiting exemplary embodiment, the subcutaneous administration method includes selecting an individual for subcutaneous administration of the pharmaceutical composition, removing or activating a partition, waiting a designated time for the dry composition and the reconstituted solution to form a reconstituted pharmaceutical composition, inserting a needle into the individual at an appropriate angle, pushing a pusher with an appropriate force, and removing the needle from the individual.

[0072] Disease being treated The compositions, methods, and devices described herein can be useful for treating diseases in individuals, such as humans. In some embodiments, the disease is one or more of pulmonary hypertension, central nervous system disorders, mitochondrial diseases, or cancer.

[0073] I. Pulmonary hypertension Pulmonary hypertension (PH) is a syndrome characterized by elevated pulmonary artery pressure. PH is hemodynamically defined as a systolic pulmonary artery pressure greater than 30 mmHg or a mean pulmonary artery pressure assessment greater than 25 mmHg. See Zaiman et al., Am. J. Respir. Cell Mol. Biol. 33:425-31 (2005).

[0074] In some embodiments of any one of the methods described herein, the disease to be treated comprises pulmonary hypertension.In some embodiments, the pulmonary hypertension is any of pulmonary arterial hypertension (PAH), idiopathic pulmonary arterial hypertension (IPAH), hereditary pulmonary arterial hypertension (HPAH), drug and toxin-induced PAH, connective tissue disease-related PAH, and congenital heart disease-related PAH.

[0075] In some embodiments, the pulmonary hypertension is severe pulmonary arterial hypertension. In some embodiments, the pulmonary hypertension is World Health Organization (WHO) functional class II, III, or IV pulmonary arterial hypertension. In some embodiments, the pulmonary hypertension is WHO functional class II pulmonary arterial hypertension. In some embodiments, the pulmonary hypertension is WHO functional class III pulmonary arterial hypertension. In some embodiments, the pulmonary hypertension is WHO functional class IV pulmonary arterial hypertension.

[0076] Central nervous system disorders Central nervous system diseases, also known as central nervous system disorders, are a range of neurological disorders that affect the structure or function of the brain or spinal cord, which collectively form the central nervous system (CNS).

[0077] In some embodiments, the CNS disorder is glioma. In some embodiments, the CNS disorder is glioblastoma. In some embodiments, the CNS disorder is epilepsy. In some embodiments, the CNS disorder is cortical dysplasia (e.g., focal cortical dysplasia). In some embodiments, the CNS disorder is selected from the group consisting of tuberous sclerosis complex, brain tumor, fragile X syndrome, Down syndrome, Rett syndrome, Alzheimer's disease, Parkinson's disease, and Huntington's disease.

[0078] In some embodiments, the CNS disorder is epilepsy. In some embodiments, the individual has undergone epilepsy surgery. In some embodiments, the individual has at least 5 seizures within 30 days after epilepsy surgery or has not been seizure-free for 1 week after epilepsy surgery. In some embodiments, the method further comprises administering to the individual an effective amount of an antiepileptic drug.

[0079] In some embodiments, the CNS disorder is glioblastoma.In some embodiments, the glioblastoma is recurrent glioblastoma.In some embodiments, the glioblastoma is newly diagnosed glioblastoma.In some embodiments, before the start of administration of nanoparticles, the individual has undergone surgical resection of newly diagnosed glioblastoma.

[0080] Mitochondrial diseases Mitochondria are organelles present in most eukaryotic cells. In addition to producing ATP, mitochondria are also involved in other cellular functions, such as cellular homeostasis, signal transduction pathways, and steroidogenesis.

[0081] Individuals with mitochondrial-associated disorders (i.e., mitochondrial diseases), including but not limited to, individuals with ataxia, nephropathy, liver disorder, metabolic disease, myopathy, neuropathy, myelopathy, encephalopathy, oxidative phosphorylation disorders, aging disorders, autism spectrum disorders, chronic inflammatory diseases, diabetes, and fatty acid oxidation disorders, can be treated by the methods disclosed herein. In some embodiments, the individual with a mitochondrial-associated disorder has a mitochondrial DNA mutation-associated disorder. In some embodiments, the individual with a mitochondrial-associated disorder has an X-chromosome mutation-associated disorder. In some embodiments, the individual with a mitochondrial-associated disorder has a nuclear DNA mutation-associated disorder. In some embodiments, the individual with a mitochondrial-associated disorder has Leigh syndrome, such as maternally inherited Leigh syndrome. In some embodiments, the Leigh syndrome is childhood-onset Leigh syndrome, juvenile-onset Leigh syndrome, or adult-onset Leigh syndrome. In some embodiments, the individual with a mitochondrial-associated disorder has MELAS syndrome. In some embodiments, the individual with a mitochondrial-associated disorder has NARP syndrome.

[0082] Individuals with metabolic disorders, including but not limited to disorders associated with cellular glucose consumption (e.g., abnormally high cellular glucose consumption in one or more tissues), insulin resistance-related disorders, hypoglycemia, autoimmune hypoglycemia, type I diabetes, type II diabetes, and metabolic syndrome, can be treated by the methods described herein.

[0083] The methods described herein can be used for any one or more of the following purposes: alleviating one or more symptoms in an individual with a mitochondrial-associated disorder, ... preventing one or more symptoms in an individual with a mitochondrial-associated disorder, treating one or more symptoms in an individual with a mitochondrial-associated disorder, ameliorating one or more symptoms in an individual with a mitochondrial-associated disorder, and delaying the onset of one or more symptoms in an individual with a mitochondrial-associated disorder.

[0084] As used herein, the term " mitochondrial-related disorder " and " mitochondrial disease " refer to any disease or disorder caused by mitochondrial dysfunction.Mitochondrial-related disorder can cause a variety of complex symptoms.Symptoms of mitochondrial-related disorder include, for example, muscle weakness, muscle spasms, seizures, food reflux, learning disabilities, hearing loss, short stature, eye muscle paralysis, diabetes, heart problems and stroke-like episodes.Symptoms of mitochondrial-related disorder can range in severity from life-threatening to barely perceptible.

[0085] Individuals with mitochondrial-associated disorders can be classified into one or more subsets of mitochondrial-associated disorders based on genotype, phenotypic expression, and / or one or more symptoms. In some embodiments, individuals with mitochondrial-associated disorders have one or more of the following: ataxia, nephropathy, liver damage, metabolic disorders, myopathy, neuropathy, myelopathy, encephalopathy, oxidative phosphorylation disorders, aging disorders, autism spectrum disorder, chronic inflammatory disease, or fatty acid oxidation disorders. In some embodiments, individuals with mitochondrial-associated disorders have one or more of the following: ataxia, nephropathy, liver damage, metabolic disorders, myopathy, neuropathy, myelopathy, encephalopathy, or oxidative phosphorylation disorders. In some embodiments, individuals with mitochondrial-associated disorders have one or more of the following: aging disorders, autism spectrum disorder, chronic inflammatory disease, diabetes, or fatty acid oxidation disorders. In some embodiments, individuals with mitochondrial-associated disorders have at least one ataxia. In some embodiments, individuals with mitochondrial-associated disorders have at least myelopathy and encephalopathy. In some embodiments, the individual with a mitochondrial-associated disorder has at least neuropathy, myelopathy, and encephalopathy. In some embodiments, the individual with a mitochondrial-associated disorder has at least myopathy and neuropathy.

[0086] Cancer treatment methods The methods described herein can be used to treat individuals with cancers that have aberrant mTOR activation in one or more genes (such as TSC1, TSC2, RPS6, PTEN, TP53, RB1, ATRX, or FAT1). In some embodiments, there are methods for treating cancer in individuals with aberrant mTOR activation in TSC2. Individuals with cancer can be selected for treatment with the methods described herein based on having aberrant mTOR activation in one or more genes (such as TSC1, TSC2, RPS6, PTEN, TP53, RB1, ATRX, or FAT1). In some embodiments, individuals are selected for treatment based on having aberrant mTOR activation in TSC2.

[0087] In some embodiments, cancer in an individual (e.g., an advanced and / or malignant cancer, e.g., a PEComa, e.g., an advanced and / or malignant cancer, e.g., a locally advanced inoperable cancer, e.g., a solid tumor) can be treated by the methods described herein, such as subcutaneously administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and a carrier protein, wherein the individual is selected for treatment based on having abnormal mTOR activation in TSC2. In some embodiments, cancer in an individual (e.g., an advanced and / or malignant cancer, e.g., a PEComa, e.g., an advanced and / or malignant cancer, e.g., a locally advanced inoperable cancer, e.g., a solid tumor) can be treated by the methods described herein, such as subcutaneously administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and a carrier protein, wherein the individual has abnormal mTOR activation in TSC2. In some embodiments, the abnormal mTOR activation in TSC2 comprises a mutation in TSC2. In some embodiments, the mutation is selected from the group consisting of a splice site mutation, a nonsense mutation, a frameshift mutation, and a missense mutation. In some embodiments, the mTOR activation abnormality in TSC2 comprises a single nucleotide polymorphism (SNV). In some embodiments, the SNV comprises a mutation selected from the group consisting of C1503T, C2743G, C5383T, C3755G, G760T, C3442T, G880A, T707C, and A4949G, or a deletion of one or more amino acids at positions 1405-1409, 1960-1970, 4999, 5002, 3521, 5208, and 5238-5255. In some embodiments, the mTOR activation abnormality in TSC2 comprises a TSC2 copy number variation. In some embodiments, the mTOR activation abnormality in TSC2 is a loss-of-function mutation. In some embodiments, the mTOR activation mutation in TSC2 comprises an abnormal expression level of TSC2. In some embodiments, the abnormal mTOR activation in TSC2 comprises an abnormal amount of activated protein encoded by TSC2.In some embodiments, the mTOR activation abnormality in TSC2 comprises loss of heterozygosity of TSC2. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the mTOR inhibitor is rapamycin or its derivative. In some embodiments, the mTOR inhibitor is rapamycin. In some embodiments, the carrier protein is albumin (such as human serum albumin). In some embodiments, the dose of the mTOR inhibitor in the composition for each administration is about 0.1 mg / m. 2 to approximately 100 mg / m 2 (e.g., about 0.1 mg / m 2 to approximately 10 mg / m 2 , about 10mg / m 2 to approximately 50 mg / m 2 , about 50mg / m 2 to approximately 100 mg / m 2 , about 75mg / m 2 to approximately 100 mg / m 2 ). In some embodiments, the method comprises subcutaneously administering a nanoparticle composition to an individual weekly for about two weeks, followed by a rest period of about one week. In some embodiments, the cancer is selected from the group consisting of pancreatic neuroendocrine tumor, endometrial cancer, breast cancer, lymphangioleiomyomatosis (LAM), prostate cancer, hepatocellular carcinoma, melanoma, renal cell carcinoma, bladder cancer, endometrial cancer, ovarian cancer, gynecological cancer, sarcoma, perivascular epithelioid cell tumor (PEComa), Hodgkin's lymphoma, and multiple myeloma. In some embodiments, the cancer is a PEComa. In some embodiments, an individual is selected for treatment based on having a TSC2 abnormality (e.g., a TSC2 mutation), regardless of the nature of the cancer. In some embodiments, the individual does not have a TSC1 abnormality (e.g., a TSC1 mutation).

[0088] In some embodiments, provided herein is a method for treating cancer (e.g., an advanced and / or malignant cancer, e.g., a PEComa, e.g., an advanced and / or malignant cancer, e.g., a locally advanced inoperable cancer, e.g., a solid tumor) in an individual, wherein the individual is selected for treatment based on having a TSC2 abnormality (e.g., a TSC2 mutation), such as subcutaneously administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and a carrier protein. In some embodiments, the present invention provides a method for treating cancer (e.g., progressive and / or malignant cancer, e.g., PEComa, e.g., progressive and / or malignant cancer, e.g., locally advanced inoperable cancer, e.g., solid tumor) in an individual, comprising subcutaneously administering an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and a carrier protein to the individual, wherein the individual is selected for treatment based on a) having a TSC2 abnormality (e.g., a TSC2 mutation), and b) having an RPS6 abnormality (e.g., an abnormal phosphorylation amount of the protein encoded by RPS6 (e.g., phosphorylation at S235, S236, S240, and / or S244 residues)). In some embodiments, provided herein is a method for treating cancer (e.g., an advanced and / or malignant cancer, e.g., a PEComa, e.g., an advanced and / or malignant cancer, e.g., a locally advanced inoperable cancer, e.g., a solid tumor) in an individual, comprising subcutaneously administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and a carrier protein, wherein the individual is selected for treatment based on a) having a TSC2 abnormality (e.g., a TSC2 mutation), and b) not having a TSC1 mutation.In some embodiments, the present invention provides a method for treating cancer (e.g., progressive and / or malignant cancer, such as PEComa, for example, progressive and / or malignant cancer, such as locally advanced inoperable cancer, for example, solid tumor) in an individual, comprising subcutaneously administering to the individual an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor and a carrier protein, wherein the individual is selected for treatment based on a) having TSC2 abnormality (e.g., TSC2 mutation), b) not having TSC1 mutation, and c) having RPS6 abnormality (e.g., abnormal phosphorylation of the protein encoded by RPS6 (e.g., phosphorylation at S235, S236, S240, and / or S244 residues)).In some embodiments, the mTOR activation abnormality in RPS6 comprises the activation state of phosphorylated S6 (pS6) (e.g., phosphorylation at S235, S236, S240, and / or S244 residues). In some embodiments, the mutation is selected from the group consisting of splice site mutation, nonsense mutation, frameshift mutation, and missense mutation. In some embodiments, the mTOR inhibitor is a limus drug. In some embodiments, the mTOR inhibitor is rapamycin or its derivative. In some embodiments, the mTOR inhibitor is rapamycin. In some embodiments, the carrier protein is albumin (such as human serum albumin). In some embodiments, the dose of the mTOR inhibitor in the composition for each administration is about 0.1 mg / m. 2 to approximately 100 mg / m 2 (e.g., about 0.1 mg / m 2 to approximately 10 mg / m 2 , about 10mg / m 2 to approximately 50 mg / m 2 , about 50mg / m 2 to approximately 100 mg / m 2 , about 75mg / m 2 to approximately 100 mg / m 2). In some embodiments, the method comprises subcutaneously administering the nanoparticle composition to the individual weekly for about two weeks, followed by a rest period of about one week. In some embodiments, the cancer is selected from the group consisting of pancreatic neuroendocrine tumor, endometrial cancer, breast cancer, lymphangioleiomyomatosis (LAM), prostate cancer, hepatocellular carcinoma, melanoma, renal cell carcinoma, bladder cancer, endometrial cancer, ovarian cancer, gynecological cancer, sarcoma, perivascular epithelioid cell tumor (PEComa), Hodgkin's lymphoma, and multiple myeloma. In some embodiments, the cancer is a PEComa. In some embodiments, an individual is selected for treatment based on having a TSC2 abnormality and an RPS6 abnormality, regardless of the nature of the cancer.

[0089] In some embodiments, provided herein are methods for treating cancer in an individual (e.g., an advanced and / or malignant cancer, e.g., a PEComa, e.g., an advanced and / or malignant cancer, e.g., a locally advanced inoperable cancer, e.g., a solid tumor), comprising subcutaneously administering to the individual a composition comprising nanoparticles comprising rapamycin or a derivative thereof and albumin, wherein the individual is selected for treatment based on a) having a TSC2 abnormality (e.g., a TSC2 mutation), and b) having an abnormal amount of phosphorylation of a protein encoded by RPS6 (e.g., phosphorylation at residues S235, S236, S240, and / or S244), wherein the dose of rapamycin or a derivative thereof in the composition for each administration is about 0.1 mg / m 2 to approximately 100 mg / m 2 (e.g., about 0.1 mg / m 2 to approximately 10 mg / m 2 , about 10mg / m 2 to approximately 25 mg / m 2 , about 25mg / m 2 to approximately 100 mg / m 2 , about 50mg / m 2 to approximately 100 mg / m 2 , about 75mg / m 2 to approximately 100 mg / m 2), where the composition is administered subcutaneously weekly for about two weeks, followed by a rest period of about one week.

[0090] In some embodiments, provided herein are methods of treating cancer (e.g., an advanced and / or malignant cancer, e.g., a PEComa, e.g., an advanced and / or malignant cancer, e.g., a locally advanced inoperable cancer, e.g., a solid tumor) in an individual, such as by subcutaneously administering to the individual a composition, such as nanoparticles, comprising rapamycin or a derivative thereof and albumin, wherein the individual is selected for treatment based on: a) having a TSC2 abnormality (e.g., a TSC2 mutation); b) not having a TSC1 mutation; and c) having an abnormal amount of phosphorylation of a protein encoded by RPS6 (e.g., phosphorylation at residues S235, S236, S240, and / or S244), wherein, for each administration, the dose of rapamycin or a derivative thereof in the composition is about 10 mg / m 2 to approximately 100 mg / m 2 (e.g., about 25 mg / m 2 to approximately 100 mg / m 2 , about 50mg / m 2 to approximately 100 mg / m 2 , about 75mg / m 2 to approximately 100 mg / m 2 ), where the composition is administered subcutaneously weekly for about two weeks, followed by a rest period of about one week.

[0091] In some embodiments, the abnormal phosphorylation level of the protein encoded by RPS6 is a positive state of phosphorylated S6 (pS6). In some embodiments, the abnormal phosphorylation level of the protein encoded by RPS6 is an increased phosphorylation level of S6 in cancer compared to control tissue. In some embodiments, the control tissue is derived from a non-cancerous tissue of an individual. In some embodiments, the control tissue is derived from the corresponding tissue of another individual who does not have cancer.

[0092] In some embodiments, provided herein is a method for treating a population of individuals with different cancers (e.g., advanced and / or malignant cancers, e.g., locally advanced inoperable cancers, e.g., solid tumors), comprising subcutaneously administering to the population of individuals an effective amount of a composition comprising nanoparticles comprising an mTOR inhibitor (e.g., rapamycin) and a carrier protein (e.g., albumin), wherein each of the individuals has a TSC2 abnormality (e.g., a TSC2 mutation). In some embodiments, the individuals do not have a TSC1 mutation.

[0093] In some embodiments, provided herein are methods of selecting an individual for treatment based on having a TSC2 mutated cancer, wherein the treatment comprises subcutaneously administering to the individual a composition comprising nanoparticles comprising rapamycin or a derivative thereof and albumin, wherein, for each administration, the dose of rapamycin or a derivative thereof in the composition is optionally about 10 mg / m 2 to approximately 100 mg / m 2 (e.g., about 25 mg / m 2 to approximately 100 mg / m 2 , about 50mg / m 2 to approximately 100 mg / m 2 , about 75mg / m 2 to approximately 100 mg / m 2 ), wherein the composition is optionally administered subcutaneously weekly for about two weeks, followed by a rest period of about one week. In some embodiments, the individual does not have a TSC1 mutation.

[0094] The cancer to be treated by the method complemented by the present application can be any cancer having one or more abnormal mTOR activation in any gene selected from the group consisting of TSC1, TSC2, TP53, RB1, ATRX, FAT1, PTEN, and RPS6. In some embodiments, the cancer has one or more abnormal mTOR activation in any one gene selected from the group consisting of TSC1, TSC2, TP53, and RPS6. In some embodiments, the cancer has at least one abnormal mTOR activation in RPS6 and at least one abnormal mTOR activation in TSC1, TSC2, or TP53. In some embodiments, the cancer has at least one abnormal mTOR activation in RPS6 and at least one abnormal mTOR activation in TSC1 or TSC2.

[0095] In some embodiments, the cancer is a solid tumor. In some embodiments, the cancer is a blood cancer.

[0096] In some embodiments, the cancer is progressive. In some embodiments, the cancer is malignant. In some embodiments, the cancer is inoperable locally advanced cancer.

[0097] In some embodiments, the cancer is selected from the group consisting of pancreatic neuroendocrine tumors, endometrial cancer, breast cancer, lymphangioleiomyomatosis (LAM), prostate cancer, hepatocellular carcinoma, melanoma, renal cell carcinoma, bladder cancer, uterine cancer, ovarian cancer, gynecological cancer, sarcoma, perivascular epithelioid cell tumor (PEComa), Hodgkin's lymphoma, and multiple myeloma.

[0098] In some embodiments, the cancer is a PEComa. In some embodiments, the cancer is an aggressive PEComa. In some embodiments, the cancer is an aggressive and malignant PEComa. In some embodiments, the PEComa is a primary uterine PEComa. In some embodiments, the PEComa is a retroperitoneal PEComa. In some embodiments, the PEComa is a primary renal PEComa. In some embodiments, the PEComa is a primary pulmonary PEComa. In some embodiments, the PEComa is a primary pelvic PEComa.

[0099] TSC2 is also known as tuberin, tuberous sclerosis complex 2 protein, protein phosphatase 1 regulatory subunit 160, TSC4, PPP1R160, and LAM. TSC2 protein functions as part of a complex with TSC1 by negatively regulating mTORC1 signaling. In some embodiments, the nucleic acid sequence of wild-type TSC2 gene is identified by GenBank accession number NC_000016.10, which is located on the front strand of chromosome 16 from nucleotide 2047936 to nucleotide 2088712 according to the human genome GRCh38.p2 assembly. Wild-type TSC2 gene comprises 42 exons. Mutations in the TSC2 gene can occur in any one or any combination of the 42 exons, or in any intron or non-coding region of the TSC2 gene.

[0100] In some embodiments, the amino acid sequence of the wild-type TSC2 protein is identified by Genbank accession number NP_000539.2. In some embodiments, the amino acid sequence of the wild-type TSC2 protein is identified by Genbank accession number NP_001070651.1. In some embodiments, the amino acid sequence of the wild-type TSC2 protein is identified by Genbank accession number NP_001107854.1.

[0101] In some embodiments, the nucleic acid sequence of the cDNA encoding the wild-type TSC2 protein is identified by Genbank accession number NM_000548.3. In some embodiments, the nucleic acid sequence of the cDNA encoding the wild-type TSC2 protein is identified by Genbank accession number NM_001077183.1. In some embodiments, the nucleic acid sequence of the cDNA encoding the wild-type TSC2 protein is identified by Genbank accession number NM_001114382.1.

[0102] In some embodiments, an individual is selected for treatment based on having mTOR activation abnormality in TSC2. In some embodiments, the mTOR activation abnormality in TSC2 comprises a mutation in TSC2. In some embodiments, the mutation is selected from the group consisting of a splice site mutation, a nonsense mutation, a frameshift mutation, and a missense mutation. In some embodiments, the mTOR activation abnormality in TSC2 comprises a single nucleotide polymorphism (SNV). In some embodiments, the SNV comprises a mutation selected from the group consisting of C1503T, C2743G, C5383T, C3755G, G760T, C3442T, G880A, T707C, A4949G, or a deletion of any one or more amino acids at positions 1405-1409, 1960-1970, 4999, 5002, 3521, 5208, 5238-5255.

[0103] In some embodiments, the mutation is a two-point mutation. In some embodiments, the mTOR activation abnormality in TSC2 is a loss-of-function mutation. In some embodiments, the mTOR activation abnormality in TSC2 comprises a homozygous deletion. In some embodiments, the mTOR activation abnormality in TSC2 comprises a TSC2 copy number variation. In some embodiments, the mTOR activation abnormality in TSC2 comprises an abnormal expression level of TSC2. In some embodiments, the mTOR activation abnormality in TSC2 comprises an abnormal activation level of the protein encoded by TSC2.

[0104] Ribosomal protein S6 (RPS6), also known as S6, is an organelle that catalyzes protein synthesis. Ribosomes are cellular organelles composed of a 40S small subunit and a 60S large subunit. Together, these subunits comprise four RNA species and approximately 80 structurally distinct proteins. This gene encodes a cytoplasmic ribosomal protein, a component of the 40S subunit. This protein belongs to the S6E family of ribosomal proteins. It is a major substrate of protein kinases in the ribosome, and a subset of five C-terminal serine residues is phosphorylated by different protein kinases. Phosphorylation is induced by various stimuli, including growth factors, tumor growth factors, and mitogens. Dephosphorylation occurs during growth arrest. This protein may contribute to the regulation of cell growth and proliferation by selectively translating specific classes of mRNA. As is typical for genes encoding ribosomal proteins, many processed pseudogenes of this gene exist scattered throughout the genome.

[0105] In some embodiments, the nucleic acid sequence of the wild-type RPS6 gene is identified by Genbank accession number NC_000009.12, which is from nucleotide 19375715 to nucleotide 19380236 on the front strand of chromosome 9 according to the GRCh38.p13 assembly of the human genome. The wild-type RPS6 gene comprises six exons. Mutations in the RPS6 gene can occur in any one or any combination of the six exons, or in any intron or non-coding region of the RPS6 gene.

[0106] In some embodiments, the amino acid sequence of the wild-type RPS6 protein is identified by Genbank accession number NM_001010.3.

[0107] In some embodiments, an individual is selected for treatment based on having mTOR activation abnormalities in RPS6. In some embodiments, the mTOR activation abnormalities in RPS6 include abnormal phosphorylation levels of the protein encoded by RPS6 (phosphorylation at S235, S236, S240, and / or S244 residues). In some embodiments, the abnormal phosphorylation levels of the protein encoded by RPS6 are positive status of phosphorylated S6 (pS6). In some embodiments, the abnormal phosphorylation levels of the protein encoded by RPS6 are increased phosphorylation of S6 in cancer compared with reference tissue. In some embodiments, the reference tissue is derived from the non-cancerous tissue of an individual. In some embodiments, the reference tissue is derived from the corresponding tissue of another individual without cancer. The phosphorylation status of S6 can be evaluated by IHC staining using an antibody that binds to phosphorylated residues in S6 (for example, an antibody that detects the endogenous amount of ribosomal protein S6 only when it is phosphorylated at Ser235 and 236). In some embodiments, the expression level of RPS6 is evaluated by immunohistochemistry. In some embodiments, the abnormal mTOR activation in RPS6 comprises an abnormal expression level of RPS6.

[0108] mTOR inhibitors In some embodiments, the method described herein comprises subcutaneous administration of nanoparticle inhibitor of mTOR inhibitor.mTOR is a serine / threonine specific protein kinase downstream of phosphatidylinositol 3-kinase (PI3K) / Akt (protein kinase B) pathway, and is an important regulator of cell survival, proliferation, stress and metabolism.Missing regulation of mTOR pathway has been found in many human cancers, and mTOR inhibition has significant inhibitory effect on tumor progression.

[0109] Mammalian target of rapamycin (mTOR), also known as functional target of rapamycin or FK506-binding protein 12-rapamycin-associated protein 1 (FRAP1), is an atypical serine / threonine protein kinase that exists in two distinct complexes: mTOR complex 1 (mTORC1) and mTOR complex 2 (mTORC2). mTORC1 is composed of mTOR, mTOR regulatory associated protein (raptor), mammalian lethal SEC13 protein 8 (MLST8), PRAS40, and DEPTOR (Kim et al. (2002). Cell 110: 163-75; Fang et al. (2001). Science 294 (5548): 1942-5). mTORC1 coordinates four major signaling inputs: nutrients (e.g., amino acids and phosphatidic acid), growth factors (e.g., insulin), energy, and stress (e.g., hypoxia and DNA damage). Amino acid availability signals to mTORC1 via the Rag and Ragulator (LAMTOR1-3) growth factor-related pathway, and hormones (e.g., insulin) signal to mTORC1 via Akt, which inactivates TSC2 and prevents mTORC1 inhibition. Alternatively, low ATP levels result in AMPK-dependent activation of TSC2 and phosphorylation of Raptor, resulting in a decrease in mTORC1 signaling proteins.

[0110] Active mTORC1 has numerous downstream biological effects, including mRNA transcription via phosphorylation of downstream targets (4E-BP1 and p70 S6 kinase), suppression of autophagy (Atg13, ULK1), ribosome biogenesis, and transcriptional activation, leading to mitochondrial metabolism or adipogenesis. Thus, mTORC1 activation promotes cell growth when conditions are favorable or promotes catabolic processes during stress or when conditions are unfavorable.

[0111] mTORC2 is composed of mTOR, rapamycin-insensitive mTOR companion (RICTOR), GβL, and mammalian stress-activated protein kinase-interacting protein 1 (mSIN1). In contrast to mTORC1, whose many upstream signals and cellular functions have been defined (see above), relatively little is known about the biology of mTORC2. mTORC2 regulates the cytoskeleton through stimulation of F-actin stress fibers, paxillin, RhoA, Rac1, Cdc42, and protein kinase Cα (PKCα). Knockdown of mTORC2 components has been observed to affect actin polymerization and disrupt cell morphology (Jacinto et al. (2004). Nat. Cell Biol. 6, 1122-1128; Sarbassov et al. (2004). Curr. Biol. 14, 1296-1302). This suggests that mTORC2 regulates the actin cytoskeleton by promoting the phosphorylation of protein kinase Cα (PKCα), the phosphorylation and relocalization of paxillin to focal adhesions, and the GTP loading of RhoA and Rac1. The molecular mechanisms by which mTORC2 regulates these processes remain unclear.

[0112] In some embodiments, the mTOR inhibitor is an inhibitor of mTORC1. In some embodiments, the mTOR inhibitor is an inhibitor of mTORC2. In some embodiments, the mTOR inhibitor is an inhibitor of both mTORC1 and mTORC2.

[0113] In some embodiments, the mTOR inhibitor is a limus drug. Examples of limus drugs include, but are not limited to, rapamycin, temsirolimus (CCI-779), everolimus (RAD001), ridaforolimus (AP-23573), deforolimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506). In some embodiments, the limus drug is selected from the group consisting of temsirolimus (CCI-779), everolimus (RAD001), ridaforolimus (AP-23573), deforolimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506). In some embodiments, the mTOR inhibitor is an mTOR kinase inhibitor such as CC-115 or CC-223.

[0114] In some embodiments, the mTOR inhibitor is rapamycin. Rapamycin is a macrolide antibiotic that inhibits the mTOR pathway by forming a complex with FKBP-12 and binding to mTORC1.

[0115] In some embodiments, the mTOR inhibitor is rapamycin (sirolimus), BEZ235 (NVP-BEZ235), everolimus (also known as RAD001, Zortress, Certican, and Afinitor), AZD8055, temsirolimus (also known as CCI-779 and Torisel), CC-115, CC-223, PI-103, Ku-0063794, INK128, AZD2014, NVP-BGT2 26, PF-04691502, CH5132799, GDC-0980 (RG7422), Torin 1, WAY-600, WYE-125132, WYE-687, GSK2126458, PF-05212384 (PKI-587), PP-121, OSI-027, Palomid 529, PP242, XL765, GSK1059615, WYE-354, and ridaforolimus (also known as deforolimus).

[0116] BEZ235 (NVP-BEZ235) is an imidazoquinoline derivative that is an mTORC1 catalytic inhibitor (Roper J, et al. PLoS One, 2011, 6(9), e25132). Everolimus is a 40-O-(2-hydroxyethyl) derivative of rapamycin that binds to the cyclophilin FKBP-12 and forms a complex with mTORC1. AZD8055 is a small molecule that inhibits mTORC1 phosphorylation (p70S6K and 4E-BP1). Temsirolimus is a small molecule that forms a complex with FK506-binding protein and inhibits mTOR activation when present in the mTORC1 complex. PI-103 is a small molecule that inhibits activation of the rapamycin-sensitive (mTORC1) complex (Knight et al. (2006) Cell. 125: 733-47). KU-0063794 is a small molecule that inhibits mTORC1 phosphorylation at Ser2448 in a dose- and time-dependent manner. INK128, AZD2014, NVP-BGT226, CH5132799, and WYE-687 are small molecule inhibitors of mTORC1. PF-04691502 inhibits mTORC1 activity. GDC-0980 is an orally bioavailable small molecule that inhibits class I PI3 kinase and TORC1. Torin1 is a potent small molecule inhibitor of mTOR. WAY-600 is a potent, ATP-competitive, and selective inhibitor of mTOR. WYE-125132 is an ATP-competitive small molecule inhibitor of mTORC1. GSK2126458 is an inhibitor of mTORC1. PKI-587 is a highly potent dual inhibitor of PI3Kα, PI3Kγ, and mTOR. PP-121 is a multi-target inhibitor of PDGFR, Hck, mTOR, VEGFR2, Src, and Abl. OSI-027 is a selective and potent dual inhibitor of mTORC1 and mTORC2 with IC50s of 22 nM and 65 nM, respectively. Palomid529 is a small molecule inhibitor of mTORC1 with no affinity for ABCB1 / ABCG2 and good brain penetration (Lin et al. (2013) Int J Cancer DOI: 10.1002 / ijc.28126 (Epub ahead of print)).PP242 is a selective mTOR inhibitor. XL765 is a dual mTOR / PI3k inhibitor targeting mTOR, p110α, p110β, p110γ, and p110δ. GSK1059615 is a novel dual inhibitor of PI3Kα, PI3Kβ, PI3Kδ, PI3Kγ, and mTOR. WYE-354 inhibits mTORC1 in HEK293 cells (0.2μM-5μM) and HUVEC cells (10nM-1μM). WYE-354 is a potent, specific, ATP-competitive inhibitor of mTOR. Deforolimus (ridaforolimus, AP23573, MK-8669) is a selective mTOR inhibitor.

[0117] Nanoparticle Composition The mTOR inhibitor nanoparticle compositions described herein include nanoparticles comprising (in various embodiments, consisting essentially of, or consisting of) an mTOR inhibitor (such as rapamycin) and albumin (such as human serum albumin).Nanoparticles of poorly water-soluble drugs (such as macrolides) are disclosed, for example, in US5916596A; US6506405B1; US6749868B1, US6537579B1, US7820788B2, and US8911786B2, as well as US2006 / 0263434A1, US2007 / 0082838A1, and WO2008 / 137148A2, each of which is incorporated herein by reference in its entirety.

[0118] Disclosed herein are compositions, such as pharmaceutical compositions, comprising nanoparticles comprising an mTOR inhibitor and albumin. The mTOR inhibitor is a drug selected from compounds that inhibit mammalian target of rapamycin (mTOR). In some embodiments, the mTOR inhibitor is rapamycin (also known as sirolimus) or an analog thereof. In some embodiments, the mTOR inhibitor is a limus drug, such as rapamycin and its analogs. Examples of limus drugs include, but are not limited to, temsirolimus (CCI-779), everolimus (RAD001), ridaforolimus (AP-23573), deforolimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506). In some embodiments, the limus drug is selected from the group consisting of temsirolimus (CCI-779), everolimus (RAD001), ridaforolimus (AP-23573), deforolimus (MK-8669), zotarolimus (ABT-578), pimecrolimus, and tacrolimus (FK-506). In some embodiments, the mTOR inhibitor is an mTOR kinase inhibitor such as CC-115 or CC-223. In some embodiments, the mTOR inhibitor is rapamycin (sirolimus), BEZ235 (NVP-BEZ235), everolimus (also known as RAD001, Zortress, Certican, and Afinitor), AZD8055, temsirolimus (also known as CCI-779 and Torisel), CC-115, CC-223, PI-103, Ku-0063794, INK128, AZD2014, NVP-BGT2 26, PF-04691502, CH5132799, GDC-0980 (RG7422), Torin 1, WAY-600, WYE-125132, WYE-687, GSK2126458, PF-05212384 (PKI-587), PP-121, OSI-027, Palomid 529, PP242, XL765, GSK1059615, WYE-354, and ridaforolimus (also known as deforolimus).

[0119] In some embodiments, the pharmaceutical composition further comprises an agent for improving the dissolution of the dry form of the composition and / or for improving the stability of the composition. In some embodiments, the additional agent comprises a sugar. The sugar can be, but is not limited to, a monosaccharide, a disaccharide, a polysaccharide, and derivatives or modifications thereof. The sugar can be, for example, mannitol, sucrose, fructose, lactose, maltose, dextrose, or trehalose. In some embodiments, the additional agent comprises glycine. Therefore, in one aspect, the present invention provides a pharmaceutical composition suitable for subcutaneous administration to an individual, comprising: a) nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin; and b) a sugar.

[0120] In some embodiments, the sugar is present in an amount effective to improve the stability of the nanoparticles in the composition compared to a nanoparticle composition that does not contain the sugar, hi some embodiments, the sugar is present in an amount effective to improve the filterability of the nanoparticles compared to a composition that does not contain the sugar.

[0121] In some embodiments, the sugar is present in an amount effective to enhance the solubility of the pharmaceutical composition, hi some embodiments, enhancing the solubility comprises enhancing the dissolution rate of the dry form of the nanoparticle composition after addition of a reconstitution solution.

[0122] In some embodiments, the sugar is present in an amount that reduces the incidence or severity of side effects after administration when the nanoparticle composition is subcutaneously administered.For example, in some embodiments, the side effect is rash, the composition is a nanoparticle that comprises an mTOR inhibitor and albumin, and the sugar is present in an amount that reduces the incidence of rash after subcutaneous administration of the nanoparticle composition.

[0123] In some embodiments, the pharmaceutical composition comprises nanoparticles comprising an mTOR inhibitor and albumin, wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 0.01:1 to about 100:1. In some embodiments, the composition comprises nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin, wherein the weight ratio of the albumin to the mTOR inhibitor (such as rapamycin) in the composition is about 18:1 or less (e.g., any of about 1:1 to about 18:1, about 2:1 to about 15:1, about 3:1 to about 12:1, about 4:1 to about 10:1, about 5:1 to about 9:1, and about 9:1). In some embodiments, the composition comprises nanoparticles comprising rapamycin or a derivative thereof and albumin, wherein the weight ratio of albumin to rapamycin or a derivative thereof in the composition is about 18:1 or less (e.g., about 1:1 to about 18:1, about 2:1 to about 15:1, about 3:1 to about 12:1, about 4:1 to about 10:1, about 5:1 to about 9:1, and about 9:1). In some embodiments, the mTOR inhibitor (such as rapamycin) is coated with albumin.

[0124] In some embodiments, the particles (e.g., nanoparticles) described herein have a mean or median diameter of about 1000, 900, 800, 700, 600, 500, 400, 300, 200, 150, 120, and 100 nm or less. In some embodiments, the mean or median diameter of the particles is about 200 nm or less. In some embodiments, the mean or median diameter of the particles is between about 20 nm and about 400 nm. In some embodiments, the mean or median diameter of the particles is between about 40 nm and about 200 nm. In some embodiments, the mean or median diameter of the nanoparticles is about 100-120 nm, e.g., about 100 nm. In some embodiments, the mean diameter of the particles is 120 nm or less. In some embodiments, the mean diameter of the particles is about 100-120 nm, e.g., about 100 nm. In some embodiments, the particles are sterile-filterable. Methods for determining average particle size are known in the art; for example, dynamic light scattering (DLS) is routinely used to determine the size of submicron particles. International Standard ISO22412 Particle Size Analysis - Dynamic Light Scattering, International Organization for Standardization (ISO) 2008 and Dynamic Light Scattering Common Terms Defined, Malvern Instruments Limited, 2011. In some embodiments, particle size is measured as the weight-weighted average particle size (Dv50) of the nanoparticles in the composition.

[0125] The compositions described herein can be stable aqueous suspensions of an mTOR inhibitor, for example, at any of the following concentrations: about 0.1 to about 200 mg / ml, about 0.1 to about 150 mg / ml, about 0.1 to about 100 mg / ml, about 0.1 to about 50 mg / ml, about 0.1 to about 20 mg / ml, about 1 to about 10 mg / ml, about 2 mg / ml to about 8 mg / ml, about 4 to about 6 mg / ml, and about 5 mg / ml. In some embodiments, the concentration of the mTOR inhibitor is at least about any of 0.2 mg / ml, 1.3 mg / ml, 1.5 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml, 40 mg / ml, 50 mg / ml, 100 mg / ml, 150 mg / ml, or 200 mg / ml.

[0126] In some embodiments, composition is a dry composition (such as freeze-dried), which can be reconstituted, resuspended or rehydrated to form a stable aqueous suspension of nanoparticles, such as mTOR inhibitor and albumin.In some embodiments, composition is a liquid (such as aqueous) composition that can be obtained by reconstituting or resuspending dry composition.In some embodiments, composition is an intermediate liquid (such as aqueous) composition that can be dried (such as freeze-dried).

[0127] In some embodiments, nanoparticles comprising an mTOR inhibitor (such as rapamycin) are associated with (e.g., coated with) albumin (such as human albumin or human serum albumin). In some embodiments, the composition comprises an mTOR inhibitor (such as rapamycin) in both nanoparticle and non-nanoparticle form (e.g., in the form of a solution or in the form of a soluble albumin / nanoparticle complex), wherein at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the mTOR inhibitor in the composition is in nanoparticle form. In some embodiments, the mTOR inhibitor (such as rapamycin) in the nanoparticles accounts for about 50%, 60%, 70%, 80%, 90%, 95%, or 99% by weight or more of the nanoparticles. In some embodiments, the nanoparticles have a non-polymeric matrix. In some embodiments, the nanoparticles comprise a core of an mTOR inhibitor (such as rapamycin) that is substantially free of polymeric materials (such as polymeric matrices).

[0128] In some embodiments, the composition comprises albumin in both the nanoparticle and non-nanoparticulate portions of the composition, wherein at least about 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the albumin in the composition is present in the non-nanoparticulate portion of the composition.

[0129] In some embodiments, the weight ratio of albumin (such as human albumin or human serum albumin) and mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is about 18:1 or less, such as about 15:1 or less, such as about 10:1. In some embodiments, the weight ratio of albumin (such as human albumin or human serum albumin) and mTOR inhibitor (such as rapamycin) in the composition is within any one of the following ranges: about 1:1 to about 18:1, about 2:1 to about 15:1, about 3:1 to about 13:1, about 4:1 to about 12:1, about 5:1 to about 10:1. In some embodiments, the weight ratio of albumin and mTOR inhibitor (such as rapamycin) in the nanoparticle portion of the composition is about 1:2, 1:3, 1:4, 1:5, 1:9, 1:10, or 1:15 or less. In some embodiments, the weight ratio of albumin (such as human albumin or human serum albumin) and mTOR inhibitor (such as rapamycin) in the composition is any one of the following: about 1:1 to about 18:1, about 1:1 to about 15:1, about 1:1 to about 12:1, about 1:1 to about 10:1, about 1:1 to about 9:1, about 1:1 to about 8:1, about 1:1 to about 7:1, about 1:1 to about 6:1, about 1:1 to about 5:1, about 1:1 to about 4:1, about 1:1 to about 3:1, about 1:1 to about 2:1, about 1:1 to about 1:1.

[0130] The nanoparticles described herein may be present in a dry formulation (e.g., a lyophilized composition) or may be present suspended in a biocompatible medium, such as a reconstitution solution. Suitable biocompatible media include, but are not limited to, water, buffered aqueous media, saline, buffered saline, suitably buffered solutions of amino acids, suitably buffered solutions of proteins, suitably buffered solutions of sugars, suitably buffered solutions of vitamins, suitably buffered solutions of synthetic polymers, lipid-containing emulsions, and the like.

[0131] In some embodiments, the pharmaceutically acceptable carrier comprises albumin (such as human albumin or human serum albumin). Albumin can be naturally occurring or synthetically prepared. In some embodiments, the albumin is human albumin or human serum albumin. In some embodiments, the albumin is recombinant albumin.

[0132] Human serum albumin (HSA) is a rHSA is a 65K highly soluble globular protein consisting of 585 amino acids. It is the most abundant protein in plasma and accounts for 70-80% of the colloid osmotic pressure of human plasma. The amino acid sequence of HSA contains a total of 17 disulfide bridges, one free thiol (Cys34), and one tryptophan (Trp214). Intravenous use of HSA solution is indicated for the prevention and treatment of hypovolemic shock (see, for example, Tullis, JAMA, 237: 355-360, 460-463, (1977) and Houser et al., Surgery, Gynecology and Obstetrics, 150: 811-816 (1980)), and in combination with exchange transfusion in the treatment of neonatal hyperbilirubinemia (see, for example, Finlayson, Seminars in Thrombosis and Hemostasis, 6, 85-120, (1980)). Other albumins, such as bovine serum albumin, are contemplated. The use of such non-human albumins may be appropriate in the context of the use of these compositions in non-human mammals, for example, in veterinary medicine (such as in the context of farm animals and agriculture). Human serum albumin (HSA) has many hydrophobic binding sites (eight in total for fatty acids, which are endogenous ligands of HSA) that bind to various drugs, particularly neutral and negatively charged hydrophobic compounds (Goodman et al., The Pharmacological Basis of Therapeutics, 9th ed., McGraw-Hill New York (1996)).Two high-affinity binding sites are presented in subdomains IIA and IIIA of HSA, which are very elongated hydrophobic pockets with charged lysine and arginine residues near the surface that serve as attachment points for polar ligand moieties (e.g., Fehske et al., Biochem. Pharmacol., 30, 687-92 (198a), Vorum, Dan. Med. Bull., 46, 379-99 (1999), Kragh-Hansen, Dan. Med. Bull., 1441, 131-40 (1990), Curry et al., Nat. Struct. Biol., 5, 827-35 (1998), Sugio et al., Protein. Eng., 12, 439-46 (1999), He et al., Nature, 358, 209-15 (199b), and Carter et al., Adv. Protein. Chem., 45, 153-203 (1994). Rapamycin and propofol have been shown to bind to HSA (e.g., Paal et al., Eur. J. Biochem., 268(7), 2187-91 (200a), Purcell et al., Biochim. Biophys. Acta, 1478(a), 61-8 (2000), Altmayer et al., Arzneimittelforschung, 45, 1053-6 (1995), and Garrido et al., Rev. Esp. Anestestiol. Reanim., 41, 308-12 (1994)).

[0133] In some embodiments, the compositions described herein are substantially free (e.g., free) of surfactants, such as Cremophor (or polyoxyethylated castor oil, e.g., Cremophor EL® (BASF)). In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., a rapamycin / albumin nanoparticle composition) is substantially free (e.g., free) of surfactants. An mTOR inhibitor nanoparticle composition (e.g., a rapamycin / albumin nanoparticle composition) is "substantially free of cremophor" or "substantially free of surfactants" if, when administered to an individual, the amount of cremophor or surfactant in the composition is not sufficient to cause one or more adverse effects in the individual. In some embodiments, the mTOR inhibitor nanoparticle composition (e.g., a rapamycin / albumin nanoparticle composition) contains less than about 20%, 15%, 10%, 7.5%, 5%, 2.5%, or 1% of any one of an organic solvent or surfactant. In some embodiments, the albumin is human albumin or human serum albumin. In some embodiments, the albumin is recombinant albumin.

[0134] The amount of albumin in the compositions described herein varies depending on other components in the composition.In some embodiments, the composition comprises albumin in an amount sufficient to stabilize the mTOR inhibitor (such as rapamycin) in aqueous suspension, for example, in the form of stable colloidal suspension (such as the stable suspension of nanoparticles).In some embodiments, the amount of albumin is such that it reduces the sedimentation rate of the mTOR inhibitor (such as rapamycin) in aqueous solvent.For particle-containing compositions, the amount of albumin also depends on the size and density of nanoparticles of mTOR inhibitor.

[0135] An mTOR inhibitor (such as rapamycin) is "stable" in an aqueous suspension if it remains suspended in the aqueous solvent (e.g., without visible precipitation or settling) for an extended period of time, such as at least about 0.1, 0.2, 0.25, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 24, 36, 48, 60, or 72 hours. The suspension is generally, but not necessarily, suitable for administration to an individual (e.g., a human). The stability of the suspension is generally (but not necessarily) evaluated at storage temperatures (e.g., room temperature (e.g., 20-25°C) or refrigerated conditions (e.g., 4°C)). For example, a suspension is stable at storage temperatures if, approximately 15 minutes after preparation of the suspension, it shows no visible aggregation or particle agglomeration when observed with an optical microscope at 1000x magnification. Stability can also be evaluated under accelerated test conditions, such as temperatures of about 40°C or higher.

[0136] In some embodiments, albumin is present in an amount sufficient to stabilize the mTOR inhibitor (such as rapamycin) in the aqueous suspension at a specific concentration. For example, the concentration of the mTOR inhibitor (such as rapamycin) in the composition is about 0.1 to about 100 mg / ml, such as about 0.1 to about 50 mg / ml, about 0.1 to about 20 mg / ml, about 1 to about 10 mg / ml, about 2 mg / ml to about 8 mg / ml, about 4 to about 6 mg / ml, or about 5 mg / ml. In some embodiments, the concentration of the mTOR inhibitor (such as rapamycin) is at least about any of 1.3 mg / ml, 1.5 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, 10 mg / ml, 15 mg / ml, 20 mg / ml, 25 mg / ml, 30 mg / ml, 40 mg / ml, and 50 mg / ml. In some embodiments, the albumin is present in an amount that avoids the use of a surfactant (such as Cremophor) such that the composition is free or substantially free of a surfactant (such as Cremophor).

[0137] In some embodiments, the composition, in liquid form, comprises about 0.1% to about 50% (w / v) albumin (e.g., about 0.5% (w / v), about 5% (w / v), about 10% (w / v), about 15% (w / v), about 20% (w / v), about 30% (w / v), about 40% (w / v), or about 50% (w / v)). In some embodiments, the composition, in liquid form, comprises about 0.5% to about 5% (w / v) albumin.

[0138] In some embodiments, the weight ratio of albumin to mTOR inhibitor (such as rapamycin) in the mTOR inhibitor nanoparticle composition is sufficient for the mTOR inhibitor to bind to or be transported by cells.The weight ratio of albumin to mTOR inhibitor (such as rapamycin) needs to be optimized for different albumin and mTOR inhibitor combinations, but generally, the weight ratio of albumin to mTOR inhibitor (such as rapamycin) (w / w) is about 0.01:1 to about 100:1, about 0.02:1 to about 50:1, about 0.05:1 to about 20:1, about 0.1:1 to about 20:1, about 1:1 to about 18:1, about 2:1 to about 15:1, about 3:1 to about 12:1, about 4:1 to about 10:1, about 5:1 to about 9:1 or about 9:1. In some embodiments, the weight ratio of albumin to mTOR inhibitor (such as rapamycin) is about any of 18:1 or less, 15:1 or less, 14:1 or less, 13:1 or less, 12:1 or less, 11:1 or less, 10:1 or less, 9:1 or less, 8:1 or less, 7:1 or less, 6:1 or less, 5:1 or less, 4:1 or less, and 3:1 or less. In some embodiments, the weight ratio of albumin (such as human albumin or human serum albumin) to mTOR inhibitor (such as rapamycin) in the composition is any of the following: about 1:1 to about 18:1, about 1:1 to about 15:1, about 1:1 to about 12:1, about 1:1 to about 10:1, about 1:1 to about 9:1, about 1:1 to about 8:1, about 1:1 to about 7:1, about 1:1 to about 6:1, about 1:1 to about 5:1, about 1:1 to about 4:1, about 1:1 to about 3:1, about 1:1 to about 2:1, about 1:1 to about 1:1.

[0139] In some embodiments, the albumin allows the composition to be administered to an individual (such as a human) without significant side effects. In some embodiments, the albumin (such as human serum albumin or human albumin) is in an amount effective to reduce one or more side effects of subcutaneous administration of an mTOR inhibitor (such as rapamycin) to a human. The term "reducing one or more side effects" of administration, such as subcutaneous administration of an mTOR inhibitor (such as rapamycin), refers to reducing, alleviating, eliminating, or avoiding one or more undesirable effects caused by the mTOR inhibitor, as well as side effects caused by the delivery of the vehicle used to deliver the mTOR inhibitor (such as a solvent that makes the limus drug suitable for injection). Such side effects include, for example, myelosuppression, neurotoxicity, hypersensitivity, inflammation, venous irritation, phlebitis, pain, dermatitis, peripheral neuropathy, neutropenic fever, anaphylactic reaction, venous thrombosis, extravasation, and combinations thereof. However, these side effects are merely exemplary, and other side effects or combinations of side effects associated with limus drugs (such as rapamycin) can be reduced.

[0140] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of 200 nm or less. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm (e.g., about 100 nm) or less. In some embodiments, the average or median diameter of the nanoparticles is about 100-120 nm, for example, about 100 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm (e.g., about 100 nm) or less. In some embodiments, the average or median diameter of the nanoparticles is about 100-120 nm, e.g., about 100 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the average or median diameter of the nanoparticles is about 10 to about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the average or median diameter of the nanoparticles is about 40 to about 120 nm.

[0141] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the composition further comprises a sugar, and wherein the nanoparticles have an average diameter of about 200 nm or less. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the composition further comprises a sugar, and wherein the nanoparticles have an average diameter of about 150 nm or less. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the composition further comprises a sugar, and wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm). In some embodiments, the average or median diameter of the nanoparticles is about 100-120 nm, for example, about 100 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the composition further comprises a sugar, and wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm). In some embodiments, the average or median diameter of the nanoparticles is about 100-120 nm, e.g., about 100 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the composition further comprises a sugar, and wherein the average or median diameter of the nanoparticles is about 10 to about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the average or median diameter of the nanoparticles is about 40 to about 120 nm. In some embodiments, the average or median diameter of the nanoparticles is about 100-120 nm, e.g., about 100 nm.

[0142] In some embodiments, the mTOR inhibitor nanoparticle composition described herein comprises nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 200 nm or less, and wherein the weight ratio of the albumin and the mTOR inhibitor in the composition is about 9: 1 or less (such as about 9: 1 or about 8: 1). In some embodiments, the mTOR inhibitor nanoparticle composition described herein comprises nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less, and wherein the weight ratio of the albumin and the mTOR inhibitor in the composition is about 9: 1 or less (such as about 9: 1 or about 8: 1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm), and wherein the weight ratio of albumin to mTOR inhibitor in the composition is about 9:1 or about 8:1. In some embodiments, the average or median diameter of the nanoparticles is about 10 nm to about 150 nm. In some embodiments, the average or median diameter of the nanoparticles is about 40 nm to about 120 nm. In some embodiments, the average or median diameter of the nanoparticles is about 100-120 nm, for example, about 100 nm.

[0143] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the composition further comprises a saccharide, wherein the nanoparticles have an average diameter of about 200 nm or less, and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the composition further comprises a saccharide, wherein the nanoparticles have an average diameter of about 150 nm or less, and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle composition described herein comprises nanoparticles comprising an mTOR inhibitor (such as rapamycin) and albumin (such as human albumin or human serum albumin), wherein the composition further comprises a saccharide, wherein the nanoparticles have an average diameter of about 150 nm, and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle composition described herein comprises nanoparticles comprising rapamycin and human albumin (such as human serum albumin), wherein the composition further comprises a saccharide, wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm), and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or about 8:1. In some embodiments, the average or median diameter of the nanoparticles is about 10 nm to about 150 nm. In some embodiments, the average or median diameter of the nanoparticles is about 40 nm to about 120 nm. In some embodiments, the nanoparticles have an average or median diameter of about 100-120 nm, eg, about 100 nm.

[0144] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 200 nm or less. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 10 nm to about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 40 nm to about 120 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin associated with (e.g., coated on) a human albumin (such as human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin associated with (e.g., coated on) a human albumin (such as human serum albumin), wherein the nanoparticles have an average diameter of about 10 nm to about 150 nm.In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin associated with (e.g., coated with) human albumin (e.g., human serum albumin), wherein the nanoparticles have an average diameter of about 40 nm to about 120 nm. In some embodiments, the average or median diameter of the nanoparticles is about 100-120 nm, e.g., about 100 nm.

[0145] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the composition further comprises a sugar. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the composition further comprises a sugar, wherein the nanoparticles have an average diameter of about 200 nm or less. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the composition further comprises a sugar, wherein the nanoparticles have an average diameter of about 150 nm or less. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the composition further comprises a sugar, and wherein the nanoparticles have an average diameter of about 10 nm to about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the composition further comprises a sugar, and wherein the nanoparticles have an average diameter of about 40 nm to about 120 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin associated with (e.g., coated on) a human albumin (such as human serum albumin), wherein the composition further comprises a sugar, and wherein the nanoparticles have an average diameter of about 150 nm (e.g., about 100 nm) or less.In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin associated with (e.g., coated on) human albumin (e.g., human serum albumin), wherein the composition further comprises a sugar, and wherein the nanoparticles have an average diameter of about 10 nm to about 150 nm. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin associated with (e.g., coated on) human albumin (e.g., human serum albumin), wherein the composition further comprises a sugar, and wherein the nanoparticles have an average diameter of about 40 nm to about 120 nm. In some embodiments, the average or median diameter of the nanoparticles is about 100-120 nm, e.g., about 100 nm.

[0146] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) albumin (such as human albumin or human serum albumin), wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 200 nm or less, and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less, and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm, and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin associated with (e.g., coated with) human albumin (such as human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm or less), and wherein the weight ratio of albumin to rapamycin in the composition is about 9:1 or about 8:1. In some embodiments, the average or median diameter of the nanoparticles is about 10 nm to about 150 nm.In some embodiments, the nanoparticles have a mean or median diameter of about 40 nm to about 120 nm, hi some embodiments, the nanoparticles have a mean or median diameter of about 100-120 nm, e.g., about 100 nm.

[0147] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the composition further comprises a saccharide, and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) an albumin (such as human albumin or human serum albumin), wherein the composition further comprises a saccharide, and wherein the nanoparticles have an average diameter of about 200 nm or less, and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (e.g., about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) albumin (such as human albumin or human serum albumin), wherein the composition further comprises a saccharide, wherein the nanoparticles have an average diameter of about 150 nm or less, and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) associated with (e.g., coated on) albumin (such as human albumin or human serum albumin), wherein the composition further comprises a saccharide, wherein the nanoparticles have an average diameter of about 150 nm or less, and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1).In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin associated with (e.g., coated on) human albumin (such as human serum albumin), wherein the composition further comprises a sugar, and wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm), wherein the weight ratio of albumin to rapamycin in the composition is about 9:1 or about 8:1. In some embodiments, the average or median diameter of the nanoparticles is about 10 nm to about 150 nm. In some embodiments, the average or median diameter of the nanoparticles is about 40 nm to about 120 nm. In some embodiments, the average or median diameter of the nanoparticles is about 100-120 nm, e.g., about 100 nm.

[0148] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 200 nm or less. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin stabilized by human albumin (such as human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm). In some embodiments, the average or median diameter of the nanoparticles is about 10 nm to about 150 nm. In some embodiments, the average or median diameter of the nanoparticles is about 40 nm to about 120 nm. In some embodiments, the average or median diameter of the nanoparticles is about 100-120 nm, for example, about 100 nm.

[0149] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further comprises a sugar. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further comprises a sugar, wherein the nanoparticles have an average diameter of about 200 nm or less. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further comprises a sugar, wherein the nanoparticles have an average diameter of about 150 nm or less. In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (e.g., rapamycin) stabilized by albumin (e.g., human albumin or human serum albumin), wherein the composition further comprises a sugar, and wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin stabilized by human albumin (e.g., human serum albumin), wherein the composition further comprises a sugar, and wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm). In some embodiments, the average or median diameter of the nanoparticles is about 10 nm to about 150 nm. In some embodiments, the average or median diameter of the nanoparticles is about 40 nm to about 120 nm. In some embodiments, the average or median diameter of the nanoparticles is about 100-120 nm, e.g., about 100 nm.

[0150] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the weight ratio of the albumin and the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 200 nm or less, and wherein the weight ratio of the albumin and the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or more, and wherein the weight ratio of the albumin and the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm, and wherein the weight ratio of the albumin and the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle composition described herein comprises nanoparticles comprising rapamycin stabilized by human albumin (such as human serum albumin), wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm), wherein the weight ratio of albumin and rapamycin in the composition is about 9: 1 or about 8: 1. In some embodiments, the average or median diameter of nanoparticles is about 10 nm to about 150 nm. In some embodiments, the average or median diameter of nanoparticles is about 40 nm to about 120 nm.In some embodiments, the mean or median diameter of the nanoparticles is about 100-120 nm, eg, about 100 nm.

[0151] In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further comprises a saccharide, and wherein the weight ratio of the albumin and the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further comprises a saccharide, and wherein the nanoparticles have an average diameter of about 200 nm or less, and wherein the weight ratio of the albumin and the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further comprises a saccharide, wherein the nanoparticles have an average diameter of about 150 nm or less, and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising an mTOR inhibitor (such as rapamycin) stabilized by albumin (such as human albumin or human serum albumin), wherein the composition further comprises a saccharide, wherein the nanoparticles have an average diameter of about 150 nm or less, and wherein the weight ratio of the albumin to the mTOR inhibitor in the composition is about 9:1 or less (such as about 9:1 or about 8:1). In some embodiments, the mTOR inhibitor nanoparticle compositions described herein comprise nanoparticles comprising rapamycin stabilized by human albumin (such as human serum albumin), wherein the composition further comprises a sugar, wherein the nanoparticles have an average diameter of about 150 nm or less (e.g., about 100 nm), and wherein the weight ratio of albumin to rapamycin in the composition is about 9:1 or about 8:1.In some embodiments, the nanoparticles have an average or median diameter of about 10 nm to about 150 nm. In some embodiments, the nanoparticles have an average or median diameter of about 40 nm to about 120 nm. In some embodiments, the nanoparticles have an average or median diameter of about 100-120 nm, e.g., about 100 nm.

[0152] In some embodiments, the mTOR inhibitor nanoparticle composition comprises nab-rapamycin. In some embodiments, the mTOR inhibitor nanoparticle composition is nab-rapamycin. Nab-rapamycin is a formulation of rapamycin stabilized with human albumin USP, which can be dispersed in a directly injectable physiological solution. The weight ratio of human albumin to rapamycin is about 8:1 to about 9:1. When dispersed in an appropriate aqueous medium, such as 0.9% sodium chloride injection or 5% dextrose injection, nab-rapamycin forms a stable colloidal suspension of rapamycin. The average particle size of the nanoparticles in the colloidal suspension is about 100 nanometers. Because HSA is readily soluble in water, nab-rapamycin can be reconstituted in a wide range of concentrations, ranging from dilute (0.1 mg / ml rapamycin or its derivatives) to concentrated (20 mg / ml rapamycin or its derivatives), for example, from about 2 mg / ml to about 8 mg / ml, or about 5 mg / ml.

[0153] The method of nanoparticle composition is known in the art.For example, the nanoparticles comprising mTOR inhibitor (such as rapamycin) and albumin (such as human serum albumin or human albumin) can be prepared under the condition of high shear force (for example, ultrasonic treatment, high-pressure homogenization, etc.).These methods are disclosed in, for example, United States Patent (USP) No. 5,916,596; No. 6,506,405; No. 6,749,868, No. 6,537,579, No. 7,820,788 and No. 8,911,786, and US Patent Publication No. 2007 / 0082838, No. 2006 / 0263434, and PCT Application No. WO08 / 137148.

[0154] Briefly, mTOR inhibitor (such as rapamycin) can be dissolved in an organic solvent, and the solution can be added to an albumin solution. The mixture is subjected to high-pressure homogenization. The organic solvent can then be removed by evaporation. The resulting dispersion can then be freeze-dried. Suitable organic solvents include, for example, ketones, esters, ethers, chlorinated solvents, and other solvents known in the art. For example, the organic solvent can be methylene chloride or chloroform / ethanol (for example, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, or 9:1 ratio).

[0155] Other Components in the mTOR Inhibitor Nanoparticle Composition The nanoparticles described herein can be present in compositions containing other drugs, carriers, excipients, diluents, or stabilizers. For example, some negatively charged components can be added to improve stability by increasing the negative zeta potential of the nanoparticles. Such negatively charged components include, but are not limited to, bile salts of bile acids such as glycocholic acid, cholic acid, chenodeoxycholic acid, taurocholic acid, glycochenodeoxycholic acid, taurochenodeoxycholic acid, lithocholic acid, ursodeoxycholic acid, dehydrocholic acid, and others; phospholipids such as lecithin (egg yolk)-based phospholipids, including the following phosphatidylcholines: palmitoyloleoylphosphatidylcholine, palmitoyllinoleoylphosphatidylcholine, stearoyllinoleoylphosphatidylcholine, stearoyloleoylphosphatidylcholine, stearoylarachidonoylphosphatidylcholine, and dipalmitoylphosphatidylcholine. Other phospholipids include L-α-dimyristoylphosphatidylcholine (DMPC), dioleoylphosphatidylcholine (DOPC), distearoylphosphatidylcholine (DSPC), hydrogenated soybean phosphatidylcholine (HSPC), and other related compounds. Negatively charged surfactants or emulsifiers, such as sodium cholesteryl sulfate, are also suitable as additives.

[0156] In some embodiments, the composition is suitable for administration to humans. In some embodiments, the composition is suitable for administration to mammals, such as domestic animals and agricultural animals, in the context of veterinary medicine. In some embodiments, the composition is suitable for administration after reconstitution.

[0157] Examples of suitable carriers, excipients, and diluents include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, saline solution, syrup, methylcellulose, methyl and propylhydroxybenzoates, talc, magnesium stearate, and mineral oil. The formulation may further include lubricating agents, wetting agents, emulsifying and suspending agents, and / or preservatives.

[0158] Preparations suitable for subcutaneous administration include aqueous and non-aqueous isotonic sterile injection solutions, which may contain antioxidants, buffers, bacteriostats, solutes to make the preparation compatible with the blood of the intended recipient, as well as aqueous and non-aqueous sterile suspensions that may contain suspending agents, solubilizers, thickeners, stabilizers, and preservatives.The preparations may be presented in unit-dose or multi-dose sealed containers such as ampoules and vials, and can also be stored in a lyophilized condition, requiring only the addition of a sterile liquid excipient, such as water, immediately before use.Extemporaneous solutions and suspensions can be prepared from sterile powders, granules, and tablets of the type described above.

[0159] In some embodiments, the composition is formulated to have a pH ranging from about 4.5 to about 9.0, such as any of the following pH ranges: from about 5.0 to about 8.0, from about 6.5 to about 7.5, and from about 6.5 to about 7.0. In some embodiments, the pH of the composition is formulated to be about 6 or higher, such as any of about 6.5, 7, or 8 (e.g., about 8). The composition can also be made isotonic with blood by the addition of a suitable viscosity-modifying agent, such as glycerol.

[0160] kit In some embodiments, the present invention provides a kit useful for various purposes, such as treating diseases in individuals.The kit of the present invention includes one or more containers containing mTOR inhibitor nanoparticle compositions (such as rapamycin / albumin nanoparticle compositions) (or unit dosage forms and / or products) suitable for subcutaneous administration, and in some embodiments, further includes a device for subcutaneously administering mTOR inhibitor nanoparticle compositions.In some embodiments, the kit further includes instructions for use according to any of the methods described herein.The kit can further include instructions for selecting individuals suitable for treatment.The instructions provided in the kit of the present invention are generally written on a label or package insert (for example, a paper sheet included in the kit), but machine-readable instructions (for example, instructions stored on a magnetic or optical storage disk) are also acceptable.

[0161] The kits of the present invention are present in suitable packaging. Suitable packaging includes, but is not limited to, vials, bottles, jars, flexible packaging (e.g., sealed Mylar® or plastic bags), etc. The kits may optionally provide additional components such as buffers and interpretable information. Thus, the present application also provides products such as vials (e.g., sealed vials), bottles, jars, flexible packaging, etc.

[0162] Instructions for use of the mTOR inhibitor nanoparticle composition typically include information such as the dose, dosing schedule, and route of administration for the intended treatment. The container may be a unit dose, bulk package (e.g., a multi-dose package), or subunit dosage form. For example, a kit may be provided containing sufficient doses of the mTOR inhibitor nanoparticle composition (e.g., a rapamycin / albumin nanoparticle composition) to provide effective treatment to an individual for an extended period of time, such as once a week, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 2 weeks, 3 weeks, 4 weeks, 6 weeks, 8 weeks, 3 months, 4 months, 5 months, 7 months, 8 months, 9 months, or more. The kit may also include multiple unit doses of the mTOR inhibitor nanoparticle composition (e.g., a rapamycin / albumin nanoparticle composition) and instructions for use, packaged in a quantity sufficient for storage and use in pharmacies, such as hospital pharmacies and compounding pharmacies.

[0163] The kit may further comprise a device comprising the mTOR inhibitor nanoparticle composition. The instructions may further comprise instructions for use of the device. [Example]

[0164] The present application may be better understood by reference to the following non-limiting examples, which are provided as exemplary embodiments of the present application. The following examples are presented to fully illustrate the embodiments, but should not be construed as limiting the broad scope of the application in any way. While several embodiments of the present application have been shown and described herein, it will be apparent that such embodiments are given by way of example only. Numerous variations, changes, and substitutions may occur to those skilled in the art without departing from the spirit and scope of the present invention. It should be understood that various modifications to the embodiments described herein may be used in carrying out the methods described herein.

[0165] Example 1: Pharmacokinetic study of ABI-009 after subcutaneous and intravenous administration in Sprague Dawley (SD) rats Female SD rats received a single dose of nab-rapamycin (ABI-009) administered subcutaneously (i.e., "SC" or "subQ") or intravenously (IV). The study design is summarized in Table 1 below. No post-dose inflammation or toxicity was observed at the subcutaneous injection site at any time point compared to the saline control (vehicle). [Table 1]

[0166] After subcutaneous or intravenous infusion of ABI-009, rapamycin concentrations in whole blood were measured at different time points. The results of the whole blood collection are summarized in Tables 2 and 3 below. [Table 2] [Table 3]

[0167] Surprisingly, subcutaneous administration improved bioavailability, as indicated by the total area under the curve (AUC), compared to intravenous administration, as summarized in Table 4 below. Subcutaneous administration of only 0.56 mg / kg ABI-009 resulted in drug exposure similar to that of IV ABI-009 (1.7 mg / kg), a dose one-third that of IV ABI-009. Furthermore, subcutaneous administration decreased the maximum concentration achieved (Cmax) and delayed the time to reach maximum concentration (time to Cmax). The peak amount of rapamycin in the blood and the AUC increased with higher subcutaneous ABI-009 doses. [Table 4]

[0168] Example 2: Biodistribution of ABI-009 after administration in rats Tissues were collected from the rats in Example 1 either 24 hours or 168 hours after administration of ABI-009 by subcutaneous (subQ) or intravenous (IV) routes (see Table 1 for study design). Rapamycin concentrations in specific rat tissues 24 or 168 hours after administration are shown in Figure 5 (bone marrow and brain), Figure 6 (heart and lung), and Figure 7 (lung and pancreas).

[0169] The subcutaneous route of administration resulted in significant distribution to all organs tested, including bone marrow, brain, heart, liver, lung, and pancreas. The organ distribution patterns were similar between subcutaneous and intravenous administration, but subcutaneous administration at a dose of 0.56 mg / kg resulted in tissue distribution similar to that of intravenous administration at a dose of 1.7 mg / kg. There was a significant decrease in rapamycin concentrations between 24 and 168 hours in well-perfused organs, including the heart, liver, lung, and pancreas. However, brain concentrations remained relatively stable at 24 and 168 hours.

[0170] To further clarify the differences in brain and blood distribution of rapamycin, further experiments were conducted in rats. Rats were administered a single subcutaneous dose of nab-rapamycin (ABI-009) at doses of 1.7 mg / kg, 9.5 mg / kg, or 17 mg / kg. Rats were sacrificed at 24, 72, and 120 hours, and whole blood and brain tissue were collected. Rapamycin concentrations were measured for each sample at each time point. As shown in Figure 5, a dose-dependent increase in brain rapamycin concentrations was observed. Surprisingly, even at the high dose of 17 mg / kg, blood rapamycin concentrations rapidly approached baseline, but brain rapamycin concentrations were well maintained throughout the entire 120 hours, even at the lowest dose. See also Figure 8.

[0171] Example 3: Sugar-containing nab-rapamycin nanoparticle formulations Nab-rapamycin (ABI-009) formulations were formulated with and without sugar, including sucrose and trehalose. The formulations were lyophilized and then reconstituted with water at various rapamycin concentrations ranging from 1 mg / ml to 40 mg / ml. The formulations were then lyophilized again and incubated at 40°C for 15 days.

[0172] After incubation, the formulations are reconstituted with water and simultaneously or subsequently assayed for albumin oligomers and polymers and reconstitution time.

[0173] Formulations that exhibit reduced albumin oligomers and polymers and / or rapid reconstitution are selected as improved formulations for subcutaneous administration.

[0174] Example 4: Toxicity study of ABI-009 after repeated subcutaneous administration in SD rats The purpose of this study was to evaluate the overall safety and local toxicity at the injection site after repeated SC injections of ABI-009 in SD rats. Clinical signs of distress were observed to determine toxicity. Skin samples from the injection site were analyzed for signs of inflammation and necrosis by histopathology.

[0175] Fifteen female Sprague Dawley (SD) rats weighing 160-180 g were used in the study. ABI-009 was dissolved in saline to prepare a stock solution (10 mg / ml), which was then further diluted in 0.9% HSA saline solution and prepared for subcutaneous administration (volume: 1.0 ml / kg).

[0176] A. Study Design The rats were divided into 5 groups of 3 rats each. The rats were weighed and administered subcutaneously every 4 days for 4 weeks (7 injections) as specified in Table 5. [Table 5] SC=subcutaneous injection

[0177] Animals were observed daily for gross clinical signs of toxicity and local injection sites were observed for reactions to subcutaneous injection.

[0178] For animals receiving ABI-009 (Groups 3, 4, and 5), whole blood samples were collected before each infusion and analyzed for traflapamycin levels.

[0179] All animals were euthanized after 4 weeks and skin samples from the local injection sites were examined by histopathology for signs of local toxicity.

[0180] B. Experimental Procedure 1. Dosing Solution Preparation Vehicle controls consisted of 0.9% saline solution and HSA in 0.9% saline solution. Based on the 9:1 albumin:rapamycin ratio of the test product ABI-009 (manufacturing lot #C345-001, Fisher lot #51394.2), the final concentration of HSA solution was 90 mg / ml. Each vial of ABI-009 (C345-001) contained 97.4 mg rapamycin and 874 mg human albumin. HSA saline was diluted from a 20% Grifols albumin stock solution (200 mg / ml).

[0181] For the ABI-009 dosing solution, a 10 mg / ml stock ABI-009 solution is first made and then diluted with the HSA-saline solution to the desired concentration for the dosing solution. A 100 mg vial of ABI-009 was dissolved in 10 ml of 0.9% saline to prepare a 10 mg / ml solution.

[0182] A 5 mg / ml ABI-009 solution was prepared by diluting 0.6 ml of the stock solution (10 mg / ml) with 0.6 ml of HSA-0.9% saline to prepare a 5.0 mg / ml solution for Group 4. A 1.7 mg / ml ABI-009 solution was prepared by diluting 0.3 ml of the ABI-009 solution from Group 4 (5.0 mg / ml) with 0.6 ml of HSA-0.9% saline to prepare a 1.7 mg / ml solution for Group 3.

[0183] 2. Administration Rats were anesthetized, weighed, and administered subcutaneous (SC) injections of ABI-009 solution, HSA solution, and saline every 4 days for 4 weeks according to Table 6 (7 injections). [Table 6]

[0184] Rats were examined daily for clinical signs of overall toxicity and local injection site responses to subcutaneous injection. To determine toxicity, clinical signs of distress were observed: piloerection, weight loss, lethargy, discharge, neurological symptoms, morbidity, redness and inflammation at the injection site, and any other signs considered abnormal in the animal's behavior. Photographs of the injection site of all rats were taken before and after SC injection.

[0185] 3. Sample Collection and Analysis For rats treated with ABI-009 (Groups 3, 4, and 5), rats were anesthetized and blood was collected for sampling into pre-chilled K2EDTA tubes before each dose (except the first dose). Whole blood was collected, stored at -80°C in labeled Eppendorf tubes, and analyzed for tracheal rapamycin levels.

[0186] All animals were euthanized at the final euthanasia time point on Day 29 (96 hours after ABI-009 administration on Day 25 of Week 4). At the final euthanasia time point, whole blood samples were collected for analysis of tracheal rapamycin concentrations. The brain, lungs, liver, heart, pancreas, and bone marrow were collected, washed with saline to remove blood, divided into two portions, snap-frozen in individually labeled tubes, and stored at -80°C. Frozen blood samples from the ABI-009-treated groups (Groups 3, 4, and 5) were shipped on dry ice to BASi. Tracheal rapamycin blood concentrations were analyzed by BASi using an LC / MS / MS method.

[0187] At the time of final euthanasia, the skin and lower dermal layers at the SC administration site were excised for histological analysis by H&E staining for signs of inflammation by histopathology. Fifteen formalin-fixed rat skin samples were routinely processed for histopathological measurements. One slide from each block was sectioned and stained with hematoxylin and eosin (H&E). Slides were evaluated by a board-certified veterinary pathologist using a light microscope. Histologic lesions were graded on a scale of 0 to 5 (0 = absent / normal, 1 = minimal, 2 = mild, 3 = moderate, 4 = marked, 5 = severe). Median scores for different groups were analyzed by t-test.

[0188] C. Results 1. Systemic toxicity Toxicity was determined by daily observations for signs of clinical distress: piloerection, weight loss, lethargy, discharge, neurological symptoms, morbidity, redness and inflammation at the injection site, and any other signs considered abnormal in the animal's behavior. Rats were normal after administration of saline, HSA, and ABI-009 in the treatment regimen (1.7-10 mg / kg, 7 doses) and showed no signs of clinical stress during the study.

[0189] There was no weight loss (<20%) during the study, and all treatment groups gained weight (Table 7). Results showed that rats tolerated subcutaneous injections of ABI-009 over the dose range of 1.7-10.0 mg / kg. [Table 7]

[0190] 2. Local toxicity Fifteen formalin-fixed rat skin samples from the SC-administered area were subjected to histopathological analysis. Histopathological findings of the skin samples showed necrosis and mixed infiltrates of inflammatory cells in perivascular areas; both lesions were observed in the subcutaneous tissue.

[0191] Necrosis was focal and characterized by loss of normal cells, neutrophil infiltration, hemorrhage, and areas of fibrin exudation, as well as adjacent fluctuating fibroproliferation. Necrosis was observed only in samples from animals treated with ABI-009 at dose levels of 5 mg / kg (Group 4, one animal with minimal necrosis) and 10 mg / kg (Group 5, all three animals with mild to marked necrosis), whereas saline (Group 1), HSA (Group 2), and ABI-009 at 1.7 mg / kg (Group 3) did not produce necrosis. See Table 8 and Figure 9. Only ABI-009 at the highest dose of 10 mg / kg showed significantly increased necrosis scores compared with the HSA group (P=0.02, t-test). [Table 8]

[0192] Mixed inflammatory cell infiltration in the subepidermal perivascular area was characterized by the infiltration and aggregation of lymphocytes, plasma cells, macrophages, and occasionally multinucleated giant cells, as well as by fluctuating numbers of neutrophils. Mixed inflammatory cell infiltration was observed in all treatment groups, with the highest median scores in animals treated with HSA (Group 2) and ABI-009 at 10 mg / kg (Group 5). For the low-dose ABI-009 injection at 1.7 mg / kg (Group 3), the median score was similar to that of the control group (Group 1) that received a saline injection. See Table 8 and Figure 9. The high mixed inflammatory cell infiltration observed in the HSA group (Group 2) compared with the saline control (P = 0.01, t-test) suggests that the local inflammation was primarily caused by the injection of the heteroprotein human serum albumin.

[0193] Representative tissue images for rats in each group are shown in Figures 10 to 14.

[0194] For the ABI-009-treated groups, there was a dose-related increase in local toxicity with increasing ABI-009 dose. At the lowest dose of 1.7 mg / kg of ABI-009, local injection site histology was similar to that of the saline control group; necrosis and subcutaneous inflammatory cell infiltration were most severe in ABI-009-treated animals at the 10 mg / kg dose level.

[0195] 3. Trough Rapamycin Blood Concentration For the ABI-009-treated groups, trough rapamycin blood samples were collected before each injection (days 5, 9, 13, 17, 21, 25, and 29) (except for the first dose on day 1) and analyzed by BASi using an LC / MS / MS method. The respective trough concentrations are shown in Table 9. Four days after SC injection, most trough rapamycin blood concentrations were consistent within the range of 2–20 ng / ml. Two samples in the ABI-009 10 mg / kg group (group 5) were clear outliers. The reason for this observation cannot be explained. However, abnormally high trough concentrations occurred only in the highest dose group of ABI-009, which also showed moderate to marked necrosis in the subcutaneous tissue, suggesting that skin lesions inhibit normal absorption of ABI-009, resulting in prolonged drug retention. [Table 9]

[0196] For each ABI-009 treatment group, trough blood rapamycin concentrations generally remained stable, so there was no significant drug accumulation over time in the study. There was a dose-dependent increase in median trough blood rapamycin concentrations with increasing ABI-009 dose. Higher trough concentrations were observed in the ABI-009 5 mg / kg group (P=0.06) and 10 mg / kg group (P=0.01) compared with the ABI-009 1.7 mg / kg group (Figure 15).

[0197] In summary, rats were normal after ABI-009 administration at the current dose regimen (1.7-10 mg / kg, seven doses), and no weight loss was observed during the study. Histopathological results showed dose-related local signs of toxicity, with moderate to marked necrosis, at the highest dose of ABI-009 (10 mg / kg). The infiltration of mixed inflammatory cells may be caused by the heteroprotein HSA. ABI-009 at 1.7 mg / kg (solution concentration 1.7 mg / ml) showed a local injection response similar to that of the saline control. There was no significant drug accumulation after repeated SC injections. Trough blood rapamycin concentrations increased with higher ABI-009 doses.

[0198] Results showed that rats tolerated multiple doses of ABI-009 systemically, ranging from 1.7-10.0 mg / kg via subcutaneous injection. Topically, ABI-009 solution at a concentration of 1.7 mg / ml was well tolerated. No adverse effects were observed with this dose.

[0199] Example 5: Antitumor activity test of nab-rapamycin A study was conducted to compare the antitumor activity of rapamycin administered by the oral route (Rapamune) with that of nab-rapamycin (ABI-009) administered by the intravenous or subcutaneous route in a human hepatocellular carcinoma xenograft mouse model.

[0200] Human cancer cells were prepared for injection into mice by thawing frozen (liquid nitrogen) SNU-398 (TSC2-deficient human liver hepatocellular carcinoma cells) obtained from ATCC® (CRL-2233®). A 75 cm 2 x 10 ... 2 The cells were dispersed in flasks and incubated at 37°C in a humidified 5% CO2 atmosphere. At 80% cell confluence, the cells were transferred to a 150cm2 flask containing fresh medium. 2 The cells were expanded in flasks and plated at 1x10 per flank of the mouse. 7 (2x10 per mouse7 ) cell targets were obtained.

[0201] Twenty athymic nude mice were housed in cages with a filter top. Cancer cells were implanted into both flanks (1 x 10 per flank) in 0.1 ml phosphate-buffered saline containing 20% Matrigel®. 7 ) was injected subcutaneously.

[0202] Treatment day 1 begins depending on the presence of tumor (tumor average ~100-150 mm 3 The animals were divided into four groups.

[0203] Group 1 included 5 mice and received saline by intravenous route twice a week for 6 weeks.

[0204] Group 2 included 5 mice and received ABI-009 at 7.5 mg / kg by intravenous route twice weekly for 6 weeks. The total rapamycin dose was 15 mg / kg / wk.

[0205] Group 3 included 5 mice and received Rapamune at 3 mg / kg orally 5 times per week for 6 weeks. The total rapamycin dose was 15 mg / kg / wk.

[0206] Group 4 included 3 mice and received ABI-009 at 7.5 mg / kg by subcutaneous route twice weekly for 6 weeks. The total rapamycin dose was 15 mg / kg / wk.

[0207] Measurements (mouse weight and tumor measurements) were taken at the designated sacrifice time and until termination 6 weeks later, or when tumors reached 2,000 mm 3 This is done three times a week (Monday, Wednesday, and Friday) until the tumor reaches its maximum volume. Signs of distress are recorded daily. The tumor is excised and stored. Blood samples are collected at the same time as tumor excision.

[0208] result: Study is ongoing. Preliminary tumor volume results (median and standard error, SEM) for each group are summarized in Table 10 below. Tumor growth inhibition (TGI) compared to saline (Group 1) and the p-value for TGI vs. saline are also listed in Table 10. Results are also summarized in Figure 16. [Table 10]

[0209] Rapamycin oral solution at 15 mg / kg / week (Group 3) produced moderate tumor growth inhibition (TGI 33.2%, P=not significant) compared with saline controls. Equivalent weekly doses of intravenous ABI-009 (Group 2) produced significantly greater TGI than oral rapamune (TGI 66.7% vs. saline controls, P=0.0016 vs. oral rapamune). However, subcutaneous ABI-009 (Group 4) produced the greatest tumor growth inhibition (TGI 89.8%, P=0.0001 vs. saline controls, P<0.0001 vs. oral rapamune).

[0210] No signs of toxicity were observed in any of the treatment groups. No significant weight loss (>10%) was observed in any of the treatment groups. Although slight weight loss was observed in the saline control group (Group 1) by Day 15, each treatment group (Groups 2-4) maintained or gained weight by Day 15. The weight results are summarized in Figure 17.

[0211] In conclusion, ABI-009 administered intravenously or subcutaneously produced significantly greater antitumor activity than comparable weekly doses of oral rapamune in a TSC2-deficient SNU-398 human hepatocellular carcinoma xenograft mouse model. Subcutaneous administration of ABI-009 was surprisingly more effective than intravenous administration. No significant toxicity or weight loss was observed in either treatment group.

Claims

1. 1. A method of treating a disease in an individual, comprising subcutaneously administering to the individual a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor and albumin, wherein the amount of mTOR inhibitor in the pharmaceutical composition is about 0.1 mg / m ２ to about 10 mg / m ２ The method is a dose of.

2. The amount of mTOR inhibitor in the pharmaceutical composition is about 1 mg / m ２ to about 10 mg / m ２ The method of claim 1, wherein the dose is

3. The amount of mTOR inhibitor in the pharmaceutical composition is about 5 mg / m ２ The method according to claim 1 or 2, wherein the dose is

4. The method according to any one of claims 1 to 3, wherein the pharmaceutical composition further comprises a sugar.

5. The method of any one of claims 1 to 4, wherein the pharmaceutical composition is administered no more than once a week.

6. 6. The method of claim 5, wherein the pharmaceutical composition is administered once a week.

7. 6. The method of claim 5, wherein the pharmaceutical composition is administered twice every three weeks.

8. The method according to any one of claims 1 to 7, wherein the disease is cancer.

9. The method according to any one of claims 1 to 7, wherein the disease is a mitochondrial disease.

10. The method of any one of claims 1 to 9, wherein the individual is a human.

11. A method for delivering an effective amount of an mTOR inhibitor to a target tissue in an individual, the method comprising subcutaneously administering a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor and albumin.

12. 12. The method of claim 11, wherein the mTOR inhibitor is a limus drug.

13. 13. The method of claim 12, wherein the limus drug is rapamycin.

14. The method according to any one of claims 11 to 13, wherein the pharmaceutical composition further comprises a sugar.

15. The amount of mTOR inhibitor in the pharmaceutical composition is about 0.1 mg / m ２ to 10 mg / m ２ The method according to any one of claims 11 to 14, wherein the dose is

16. The method of any one of claims 11 to 15, wherein the target tissue is brain tissue of an individual.

17. A pharmaceutical composition suitable for subcutaneous administration to an individual, comprising: a) nanoparticles comprising an mTOR inhibitor and albumin; and b) a sugar.

18. 18. The pharmaceutical composition of claim 17, wherein the saccharide is selected from the group consisting of alginate, starch, lactose, pullulan, hyaluronic acid, chitosan, glucose, galactose, mannose, N-acetylglucosamine, sucrose, N-acetyl-D-galactosamine, maltose, or trehalose.

19. 19. The pharmaceutical composition of claim 18, wherein the sugar is sucrose.

20. 19. The pharmaceutical composition of claim 18, wherein the sugar is trehalose.

21. The pharmaceutical composition of any one of claims 17 to 20, wherein the concentration of the mTOR inhibitor in the pharmaceutical composition is at least about 5 mg / ml.

22. The pharmaceutical composition of any one of claims 17 to 21, wherein the concentration of the mTOR inhibitor in the pharmaceutical composition is at least about 50 mg / ml.

23. The method of any one of claims 1 to 16, or the pharmaceutical composition of any one of claims 17 to 22, wherein the average diameter of the nanoparticles in the pharmaceutical composition is about 120 nm or less.

24. The method of any one of claims 1 to 16 or the pharmaceutical composition of any one of claims 17 to 23, wherein the nanoparticles comprise an mTOR inhibitor coated with albumin.

25. The method of any one of claims 1 to 16, or the pharmaceutical composition of any one of claims 17 to 24, wherein the albumin is human albumin.

26. The method of any one of claims 1 to 16, or the pharmaceutical composition of any one of claims 17 to 25, wherein the mTOR inhibitor is a limus drug.

27. 27. The method or pharmaceutical composition of claim 26, wherein the mTOR inhibitor is rapamycin.

28. 1. A device for subcutaneously administering to an individual a pharmaceutical composition comprising nanoparticles comprising an mTOR inhibitor and albumin, the device comprising: a) a drug chamber containing the pharmaceutical composition in dry form, and a solution chamber containing a reconstitution solution; b) A removable partition separating the drug chamber from the solution chamber wherein removal of the partition causes mixing of the dried pharmaceutical composition with a reconstitution solution, thereby forming a reconstituted pharmaceutical composition.

29. 29. The device of claim 28, wherein the device is a syringe comprising an injection needle affixed to the end of the syringe and a pusher capable of expelling the reconstituted pharmaceutical composition from the syringe.

30. 30. The device of claim 28 or 29, wherein the pharmaceutical composition further comprises a sugar.

31. The device of any one of claims 28 to 30, wherein the mTOR inhibitor is a limus drug.

32. The device of any one of claims 28 to 31, wherein the mTOR inhibitor is rapamycin.

33. A kit comprising a device according to any one of claims 28 to 32 for use in the treatment of a disease.

34. 34. The kit of claim 33, further comprising instructions for using the kit to treat cancer or a mitochondrial disease.

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